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Morphological and functional plasticity of olfactory ensheathing cells

Article in Journal of Neurocytology · April 2005

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Journal of Neurocytology 34, 65–80 (2005)

Morphological and functional plasticity of olfactory

ensheathing cells
A D E L E J . V I N C E N T 1 , A D R I A N K . W E S T 2 a n d M E N G I N N C H U A H 1, ∗
1
NeuroRepair Group, Discipline of Anatomy and Physiology, Private Bag 24, University of Tasmania Hobart, Tasmania, Australia 7001;
2
NeuroRepair Group, Discipline of Biochemistry, Private Bag 58, University of Tasmania, Hobart, Tasmania, Australia 7001
Inn.Chuah@utas.edu.au

Received 2 January 2005; revised 4 April 20005 ; accepted 8 July 2005

Abstract
In the primary olfactory pathway, olfactory ensheathing cells (OECs) extend processes to envelop bundles of olfactory axons
as they course towards their termination in the olfactory bulb. The expression of growth-promoting adhesion and extracellular
matrix molecules by OECs, and their spatially close association with olfactory axons are consistent with OECs being involved
in promoting and guiding olfactory axon growth. Because of this, OECs have been employed as a possible tool for inducing
axonal regeneration in the injured adult CNS, resulting in significant functional recovery in some animal models and promising
outcomes from early clinical applications. However, fundamental aspects of OEC biology remain unclear. This brief review
discusses some of the experimental data that have resulted in conflicting views with regard to the identity of OECs. We present
here recent findings which support the notion of OECs as a single but malleable phenotype which demonstrate extensive
morphological and functional plasticity depending on the environmental stimuli. The review includes a discussion of the
normal functional role of OECs in the developing primary olfactory pathway as well as their interaction with regenerating
axons and reactive astrocytes in the novel environment of the injured CNS. The use of OECs to induce repair in the injured
nervous system reflects the functional plasticity of these cells. Finally, we will explore the possibility that recent microarray
data could point to OECs assuming an innate immune function or playing a role in modulating neuroinflammation.

Introduction
The primary olfactory pathway consists of olfactory
neurons in the nasal cavity whose axons project through
the cribriform plate into the CNS to synapse with mitral
and tufted cells in the olfactory bulb. Along the course
of the unmyelinated olfactory nerves, they are accom-
panied by glial cells, widely known as olfactory en-
sheathing cells (OECs), which extend cytoplasmic pro-
cesses to compartmentalise the small olfactory axons
(0.1–0.5 µm) into fascicles (Doucette et al., 1983).
The designation of OECs into a preexisting class of
glial cell has been a contentious issue since their discov-
ery (Barnett, 2004; Barnett & Chang, 2004; Boyd et al.,
2003; Doucette & Devon, 1993; Raisman, 2001; Ramón-
Cueto & Valverde, 1995; Wewetzer et al., 2002). This dis-
pute stems partly from the lack of a definitive molecu-
lar identity for OECs. Initially OECs were called olfac-
tory nerve Schwann cells (SCs), suggesting that save
for their location, OECs possess properties typical of
peripheral nerve glia (Barber & Lindsay, 1982; Chuah
& Au, 1991; Cuschieri & Bannister, 1975; Gong et al.,
1994). However, it soon became apparent that OECs
assume astrocyte (AC)-like functions in the CNS as ex-
emplified by their formation of the glia limitans in the
olfactory bulb (Doucette, 1991); thus it was proposed
that they could belong to the astrocyte family. Even so,
OECs were given an independent name, ‘ensheathing
cells’, in recognition of their potential to form a new
class of glia (Doucette, 1990). The notion of OECs con-
stituting a distinct class of glia has been reinforced in re-
cent years in in vivo regeneration studies (Lakatos et al.,
2003) and tissue culture (Lakatos et al., 2000) in which
they have demonstrated clear differences with SCs in
terms of cellular interaction with astrocytes and CNS
tissues.
In recent years, debate has extended to the question
of whether OECs should be classified as belonging ei-
ther to a single population or to two subpopulations
based on their morphology and antigenic profile. This
brief review will discuss findings from various in vivo
and in vitro studies which have contributed to this de-
bate, including the morphological plasticity of OECs.
This is followed by a brief discussion of the normal

∗ To whom correspondence should be addressed.

0300–4864 
C 2005 Springer Science + Business Media, Inc.
66 V I N C E N T, W E S T and C H U A H

functional role of OECs with regard to developing olfac-
tory nerves, and their physical interaction with regener-
ating axons and reactive astrocytes in the injured CNS.
Lastly we will raise the possibility that OECs could have
other possible functional roles not normally associated
with these cells, e.g. as a member of the innate immune
system.
Antigenic characteristics of OECs in vivo
Early suggestions that antigenically distinct subpopu-
lations of OECs exist in the olfactory system were based
on the variable intensity of immunohistochemical stain-
ing of the structural protein glial fibrillary acidic protein
(GFAP) in peripheral olfactory nerves (Barber & Lind-
say, 1982; Pixley, 1992). It is now clear that OECs, both in
the peripheral olfactory nerves and the olfactory nerve
layer (ONL) of the bulb, express a number of antigens
that can be detected immunohistochemically (Fig. 1;
note that for the purpose of this review, the following
discussion will only cover some of the molecules listed
in this figure). For example, research has demonstrated
the differential expression of OEC markers in the ONL.
The general consensus appears to be that all OECs ex-
press S100β and weak GFAP (and until recently, O4),
while (i) those in the outer ONL are weakly positive for
the low-affinity neurotrophin receptor p75NTR and the
embryonic neural cell adhesion molecule (E-NCAM),
and (ii) those in the inner ONL are neuropeptide Y
(NPY)-positive, but lack p75NTR and E-NCAM (Astic
et al., 1998; Barnett et al., 1993; Franceschini & Barnett,
1996; Ubink et al., 1994; Valverde et al., 1992).

Fig. 1. A diagram representing the adult rat primary olfactory system. Olfactory ensheathing cells (OEC, pink areas) ensheath
bundles of olfactory receptor axons (olfactory nerves, ON) along their course through the lamina propria in the PNS. Olfactory
nerves and their accompanying OECs cross through the cribriform plate of the skull into the CNS and surround the olfactory
bulb to form the olfactory nerve layer. The olfactory nerve layer is comprised of outer and inner layers which represent zones
of axon-sorting and molecular expression by OECs. Axons then extend unaccompanied by OECs into glomeruli to synapse on
target cells. Olfactory ensheathing cells also contribute to the formation of the glial limitans (GL). Listed below are some of the
molecules produced by OECs in the lamina propria and olfactory nerve layer. BG, Bowman’s glands; BV, blood vessel; ORN,
olfactory receptor neuron.
Morphological and functional plasticity of olfactory ensheathing cells
S100β is one of a family of calcium-binding pro-
teins which are involved in the regulation of numer-
ous intracellular processes including protein phospho-
rylation, proliferation and differentiation, and is ex-
pressed widely being present in SCs, ACs and oligo-
dendrocytes (Donato, 1999). GFAP belongs to the class
of intermediate-sized cytoskeletal proteins and is com-
monly accepted as a marker for astrocytes. In rats
S100β and GFAP appear first in the peripheral olfac-
tory nerves at E14 and E15 respectively, and advance to
the ONL just prior to birth (Astic et al., 1998; Valverde
et al., 1992). A gradient also exists between the inner
and outer sublaminae of the ONL; both markers are
restricted initially to the outer ONL and appear later
in the inner ONL, around birth for S100β and increas-
ing slowly from P7 to P20 for GFAP (Astic et al., 1998;
Valverde et al., 1992). This delay in glial maturation
may allow prolonged neuroplasticity during the de-
velopmental period of major innervation of the bulb,
a phenomenon also observed in AC precursors (As-
tic et al., 1998). The adult expression pattern of these
markers is generally thought to be uniform throughout
the ONL, with more intense immunostaining for S100β
than GFAP (Au et al., 2002; Franceschini & Barnett, 1996;
Ubink et al., 1994). However, some of this GFAP stain-
ing may be due to the presence of interfascicular ACs
(Barnett et al., 1993) which infiltrate the ONL during
development (Doucette, 1984) or it may be due to the
extended processes of periglomerular ACs (Astic et al.,
1998; Treloar et al., 1999).
The expression of p75NTR , E-NCAM and O4 is promi-
nent in OECs throughout the olfactory system during
development and later decreases to very low levels in
the adult rat (Barnett et al., 1993; Franceschini & Bar-
nett, 1996; Gong et al., 1994; Miragall & Dermietzel,
1992; Miragall et al., 1988; Miragall et al., 1989; Turner
& Pérez-Polo, 1992; Vickland et al., 1991). Interestingly,
p75NTR has been reported to be entirely absent from
the innermost area of the ONL (Au et al., 2002; Gong
et al., 1994; Treloar et al., 1999; Ubink & Hökfelt, 2000;
Vickland et al., 1991). However, some confusion exists;
for example, OECs were attributed to strong E-NCAM
staining in the outer ONL of adult rats (Franceschini &
Barnett, 1996), although immunoelectron microscopy
has shown that, at least in mice, E-NCAM is expressed
by both OECs and axons during development and is
later absent in OECs (Miragall & Dermietzel, 1992; Mi-
ragall et al., 1988). Similarly, strong p75NTR staining was
reported to be present in OECs in the outer ONL of adult
mice (Au et al., 2002), although the localisation of these
molecules on glial or axonal membranes was not con-
clusively demonstrated. Furthermore, O4, which was
reported to be co-expressed with p75NTR (Barnett et al.,
1993), was shown by others to be present throughout
the ONL and in the glomerular layer (Franceschini &
Barnett, 1996). In fact, recent work has revealed that
OECs do not express O4, but are immunopositive in
67
freshly dissociated culture because of O4-positive ax-
onal membrane fragments which adhere to the surface
of OECs and are gradually phagocytosed (Wewetzer
et al., 2005).
Neuropeptide Y displays the third expression pattern
of OEC markers. It is thought to play a role in olfactory
neuronal differentiation or axonal growth because it is
expressed in OECs in the developing ONL at E15 prior
to the emergence of other laminae in the olfactory bulb
(Ubink & Hökfelt, 2000). In adulthood it clearly delin-
eates the inner ONL from the glomerular layer which
does not stain for NPY, with OECs in the peripheral
part of the ONL showing only weak staining (Au et al.,
2002; Ubink et al., 1994; Ubink & Hökfelt, 2000).
Because of the apparent difference in staining pat-
tern for NPY, E-NCAM and p75NTR between the inner
and outer layers, it was thought that two subpopula-
tions of OECs exist in the adult ONL (Au et al., 2002;
Barnett & Chang, 2004; Franceschini & Barnett, 1996;
Ubink & Hökfelt, 2000). However, the exact definition
of these subpopulations is questionable due to variabil-
ity among studies resulting from factors such as, differ-
ential regulation of antigenic expression during devel-
opment and in adulthood, the use of different strains
or species of organisms and a variety of antisera, and
the difficulty in identifying labelled structures by light
microscopy. This latter problem is especially difficult
when immunostaining is detected in olfactory nerves
in which molecules may be expressed by both OECs
or olfactory axons, such as for the well-established
OEC markers E-NCAM, p75NTR and O4. Although re-
searchers have speculated and proposed possible func-
tions of markers such as p75NTR and NPY in OECs, un-
equivocal direct evidence that support these assertions
is still lacking. These uncertainties underscore the need
for a greater understanding of OEC biology, before we
can establish a robust definition for OECs.
Despite these uncertainties, all OECs are ultrastruc-
turally similar and are derived during ontogeny from
precursors originating from the olfactory epithelium
(Chuah & Au, 1991; Doucette, 1989; Valverde et al.,
1992). As the primary olfactory pathway differentiates,
OECs become distributed along the olfactory nerves in
the lamina propria as well as within the ONL of the ol-
factory bulb. Olfactory ensheathing cells can be readily
distinguished from other glia by their electron dense cy-
toplasm, scattered intermediate filaments and irregular
nuclei (Chuah & Au, 1993; Cuschieri & Bannister, 1975;
Doucette, 1984; Doucette, 1989; Valverde et al., 1992).
They ensheath olfactory axons, form the glial limitans
of the olfactory bulb and are present throughout the
full depth of the ONL (Au et al., 2002; Doucette, 1984;
Doucette, 1989; Valverde et al., 1992). They are clearly
distinct from interfascicular ACs, which have an elec-
tron lucent cytoplasm, multiple bundled intermediate
filaments and are never observed to ensheath axons
(Doucette, 1984, 1993).
68
Antigenic characteristics of OECs in vitro
The notion that there are two subpopulations of OECs
in vivo has been tested on cultured cells based on the as-
sumption that this diversity should be reflected in vitro.
Indeed, many studies have demonstrated antigenic het-
erogeneity in cultured OECs, yet comparison between
these studies is difficult because each research group
has developed unique culturing strategies and different
criteria for defining OECs (Table 1). Olfactory ensheath-
ing cells have been cultured from olfactory mucosa and
bulb of both neonatal and adult rats and mice of various
strains, using a variety of media, substrates, mitogens,
time in culture, number of passages and purification
techniques (Alexander et al., 2002; Barber & Lindsay,
1982; Chuah & Au, 1993; Pixley, 1992). Although a num-
ber of studies have indicated that OEC heterogeneity
exists amongst olfactory bulb-derived OECs (Alexan-
der et al., 2002; Barnett et al., 1993; Franceschini & Bar-
nett, 1996) and to some extent amongst peripherally-
derived OECs (Barber & Lindsay, 1982; Pixley, 1992),
there is no experimental data to support a clear anti-
genic or functional distinction between these two spa-
tially different sources of OECs (Au & Roskams, 2003;
Jani & Raisman, 2004; Kumar et al., 2005).
Peripherally located OECs have been observed to dis-
play varied intensities of GFAP immunostaining both in
vivo and in vitro (Barber & Lindsay, 1982; Pixley, 1992).
The majority of GFAP-positive cells from cultures of
neonatal and adult rat olfactory mucosa were diffusely
immunostained and had spindly morphology, while a
smaller number had intense filamentous staining and
a flat morphology (Barber & Lindsay, 1982). In another
study, OECs cultured from neonatal rat olfactory mu-
cosa were similarly described and were further classi-
fied based on their resemblance to SCs or to ACs (Pix-
ley, 1992). Schwann cell-like OECs were described as
p75NTR -, S100β- and GFAP-positive (the latter generally
weak) with primarily process-bearing morphologies,
whereas AC-like OECs lacked p75NTR but were GFAP-
positive (mostly fibrous or bright) and gained S100β in
culture, with various flattened morphologies (Pixley,
1992). The AC-like OECs comprised only a small per-
centage of the total GFAP-positive population (Barber
& Lindsay, 1982; Pixley, 1992) and were nearly elimi-
nated from the cultures when care was taken not to har-
vest central tissue (Pixley, 1992), suggesting that some
of these AC-like OECs could in fact have been bona fide
ACs.
The categorisation of OECs into SC-like and AC-like
subpopulations was similarly adopted by others work-
ing on OECs cultured from olfactory bulbs (Frances-
chini & Barnett, 1996; Li et al., 1998). Olfactory ensheath-
ing cells derived from neonatal rat olfactory bulbs
by fluorescence-activated cell sorting of O4-positive
cells (Alexander et al., 2002; Barnett et al., 1993) were
shown to contain subpopulations that displayed sim-
V I N C E N T, W E S T and C H U A H
ilar antigenic and morphological phenotypes to those
described previously (Pixley, 1992). These subpopula-
tions were additionally defined by their E-NCAM ex-
pression, where SC-like OECs were E-NCAM-negative
and AC-like OECs were E-NCAM-positive (Frances-
chini & Barnett, 1996). However, the AC-like subpopu-
lation was defined differently in cultures derived from
the ONL and glomerular layers of adult rat olfactory
bulb (Li et al., 1998). In this case, AC-like cells were
described as flat and immunopositive for vimentin and
fibronectin, with some cells also GFAP-positive (Li et al.,
1998). It was conceded that this subpopulation was
most likely to have been fibroblasts which may be as-
sociated with the outer meningeal surface of the ONL
(Li et al., 1998; Raisman, 2001), although the portion ex-
pressing GFAP would probably have been ACs given
that the culture was not purified and included adult
glomerular tissue.
Hence, there are a number of inconsistencies with
the interpretation that cultured OECs are comprised
of two subpopulations. (i) The identity of the ‘AC-
like’ subpopulation in each of these studies is question-
able. This subpopulation undoubtedly contained an el-
ement of contaminating ACs and fibroblasts (Li et al.,
1998; Pixley, 1992; Raisman, 2001) and possibly other
unidentified cells types (Barber & Lindsay, 1982). (ii)
The E-NCAM-positive subpopulation may also have
contained periglomerular ACs because these cells ex-
press E-NCAM in vivo (Franceschini & Barnett, 1996;
Miragall & Dermietzel, 1992). (iii) In the absence of lo-
calisation on an ultrastructural level or confirmation
by additional techniques, the expression of putative
marker molecules based on light microscopy could be
an artifact in some instances. For example, it has been
discovered recently that positive immunoreactivity for
O4 on OECs is due to the adhesion of O4-positive ax-
onal membrane to OECs (Wewetzer et al., 2005). Hence,
the categorisation of OECs into two subpopulations,
based solely on immunohistochemical staining, needs
to be interpreted with caution. (iv) Some of the puta-
tive marker molecules used to define subpopulations
of OECs may not be a consistent feature but demon-
strates variability depending on OEC interaction with
other tissues. For example, the expression of p75NTR
by OECs is regulated by axonal contact (Ramón-Cueto
et al., 1993; Vickland et al., 1991; Wewetzer et al., 2005).
In the adult ONL, OECs down-regulate expression of
p75NTR on parts of their membranes which are in direct
contact with olfactory axons while parts which face the
pia mater continue to express p75NTR (Vickland et al.,
1991). This regulation has also been demonstrated in
vitro by two methods. In coculture with olfactory neu-
rons, p75NTR -positive OECs ensheathed neurites and
lost p75NTR on their membranes where they were in di-
rect contact with the neurites (Ramón-Cueto et al., 1993).
In freshly dissociated culture, most OECs were p75NTR -
negative and had fragments of axonal membrane
Table 1. Strategies of purification and criteria for defining OECs in vitro prior to transplantation into injured CNS tissue.

Research group Animal Tissue source Purification strategy Criteria or description Degree of purity
∗
Ramón-Cueto et al. (1998, 2000), adult Wistar rat OB ; ONL + GL p75+ immunopurification p75+, p75/S100+ ∼ 98%
Ramón-Cueto and Nieto-
Sampedro (1994), Gudiño-
Cabrera and Nieto-Sampedro
(1996), Pascual et al. (2002),
Verdú et al. (1999),
Barnett et al. (2000) adult human OB p75+ immunopurification p75+ high‡
Polentes et al. (2004) adult Sprague-Dawley rat OB Thy1.1/IB4–immunopurification p75+ 75 ± 12%
Gomez et al. (2003) adult Wistar rat ONL + GL Thy1.1/IB4–immunopurification p75/GFAP+ ∼ 90%
Riddell et al. (2004) postnatal§ Fischer rat OB O4+/GalC–immunopurification p75+ highly variable
Smith et al. (2001) postnatal Fischer rat OB O4+/GalC–immunopurification p75+ >95% , predominantly p75+
Ramer et al. (2004) postnatal mouse OM Thy1.1–immunopurification p75/S100/GFAP+ >98%
(GFP transgenic)
Chuah et al. (2003) postnatal Wistar rat ONL + LP mitotic inhibitor p75/GFAP+ 92%
Smith et al. (2002) adult dog ONL shaking, serial passage p75/GFAP/O4+ 92%
Nash et al. (2002) adult Sprague-Dawley rat ON + ONL + GL differential adhesion p75/GFAP+ ∼ 93%
Lakatos et al. (2003) postnatal Fischer rat rostral ONL mitotic inhibitor, shaking, p75+ p75/GFAP+ purified 94% p75/GFAP+,
immunopurification OR unpurified ∼ 75% p75/GFAP+
unpurified (except shaking)
Li et al. (1998, 2003a) adult inbred AS rat ONL + GL none p75/S100+ OECs, ∼ 50% of each
vimentin/fibronectin+ ONFs
Kato et al. (2000) adult human, ON none GFAP/O4/vimentin+, >90% GFAP/vimentin+,
adult Wistar rat S100/GalC/A2B5– ∼ 80% O4+
Radtke et al. (2003) adult pig (HT transgenic) ONL + GL none p75/GFAP/S100+ ∼ 98% p75+
Morphological and functional plasticity of olfactory ensheathing cells

Smale et al. (1996); Boyd et al. (2004) embryonic Wistar rat ONL none S100/p75+ most S100+, part p75+
Pérez-Bouza et al. (1998) adult Lewis rat ON none n.d.# n.d.
Taylor et al. (2001) n.s. n.s. n.s. n.s. n.s.
Boruch et al. (2001) postnatal Sprague-Dawley rat OB n/a-clonal cell line nOEC¶ p75+ 100%
Imaizumi et al. (1998) postnatal Wistar rat ONL n/a-acute dissociated cell n/a n/a
transplantation
Lu et al. (2002) adult Sprague-Dawley rat LP n/a-acute tissue graft n/a n/a
Guntinas-Lichius et al. (2002) adult inbred Lewis rat OM n/a-acute tissue graft n/a n/a

∗ Abbreviations: OB: olfactory bulb, ONL: olfactory nerve layer, GL: glomerular layer, OM: olfactory mucosa, LP: lamina propria, ON: olfactory nerves, GFP: green fluorescent protein,
HT: H-transferase, OECs: olfactory ensheathing cells, ONFs: olfactory nerve fibroblasts, n.s.: not stated, n/a: not applicable, n.d.: not determined.
§up to 7 days after birth; ‡ p75 was rapidly lost in culture and transplanted cell suspensions contained 30% or 45% p75+ cells; ¶ Goodman et al., 1993; #not determined prior to transplantation
but most cells p75/nestin/vimentin+ at 6h post-transplantation.
69
70
adhering to their surface (Wewetzer et al., 2005). These
cells became p75NTR -positive after several days when
the axon fragments had been phagocytosed (Wewetzer
et al., 2005), while the small population of OECs that
did not have adherent axon fragments were p75NTR -
positive from the outset (Wewetzer et al., 2005). (v)
Finally, the identity of the OEC subpopulations de-
scribed in vitro does not match the description of those
in vivo. Cultured OECs are defined by expression of
either p75NTR or E-NCAM, whereas OECs in vivo are
thought to express both p75NTR and E-NCAM or other-
wise to express NPY alone. It would be interesting to in-
vestigate whether cultured OECs express both p75NTR
and NPY, or whether a NPY-positive/p75NTR -negative
subpopulation is present and persistent under various
culture conditions.
Franceschini and Barnett (1996) reported that the
phenotypes displayed by the two putative subpopu-
lations of OECs were also highly dependent on the
culture conditions. For example, the expression of
E-NCAM and O4 was lost after 7 days in serum-
containing medium and the subpopulations were con-
sequently defined in serum-free conditions in which
they were more distinguishable. However, it should
be noted that E-NCAM and O4 may be attributed to
fragments of axonal membrane and their “disappear-
ance’’ may be a consequence of phagocytosis by OECs
(Wewetzer et al., 2005), a process which would have
progressed more slowly in serum-free conditions (Mor-
ris et al., 2003). Cells adopted divergent morpholo-
gies when long-term cultures were switched between
different types of media (Alexander et al., 2002). Un-
der these conditions E-NCAM expression oscillated
markedly but p75NTR and GFAP remained constant
(Alexander et al., 2002). Increasing cell density also
correlated with increasing expression of p75NTR and
GFAP in serum-containing medium, although p75NTR
decreased at later passages while E-NCAM increased
(Sonigra et al., 1999).
These findings suggest that OECs have a malleable
phenotype that is dependent on environmental stimuli
(Au & Roskams, 2003; Chuah & West, 2002; Doucette,
1995). A recent study by our laboratory showed that
OECs derived from the olfactory mucosa and ONL
underwent rapid and reversible change between flat
morphology in the presence of serum and process-
bearing morphology in the absence of serum (Vin-
cent et al., 2003). Purified OECs were 87–90% flat in
serum-containing medium and became 78–84% bipo-
lar or multipolar in serum-free medium within 24 h of
the change in conditions. Rapid morphological change
in OECs was mediated by the small guanosine triphos-
phatase RhoA signalling cascade which induces equiv-
alent changes in AC morphology and is important in
cytoskeletal organisation in a variety of other cells (Hall,
1998; Ridley & Hall, 1992; Vincent et al., 2003; Yan et al.,
2003). RhoA is activated by endothelin-1 and lysophos-
V I N C E N T, W E S T and C H U A H
phatidic acid which induce OECs to adopt a flat mor-
phology probably via downstream assembly of actin
stress fibres and associated focal adhesions (Safavi-
Abbasi et al., 2001; Suidan et al., 1997). Inactivation of
RhoA by dBcAMP leads to the disassembly of these cy-
toskeletal structures causing rapid cytoplasmic retrac-
tion (Goldman & Abramson, 1990; Ramakers & Moole-
naar, 1998; Suidan et al., 1997) and the formation of
a stellate morphology in OECs. The intermediate fil-
ament protein GFAP appears to be unaffected by this
morphological change in OECs, which does not require
de novo synthesis or degradation of cytoskeletal proteins
(Safavi-Abbasi et al., 2001; Vincent et al., 2003). More
recent studies have captured the highly dynamic na-
ture of OEC morphology in culture using time-lapse
microscopy; individual OECs were observed to switch
from one morphology and back again within an hour
(Van Den Pol & Santarelli, 2003).
From an assessment of all the data now available re-
garding OEC biology, it would appear that the differ-
ential expression of markers by OECs, both in vivo and
in vitro, is more likely a reflection of their functional
plasticity rather than their existence as discrete, inex-
changable subpopulations. The stongest data in sup-
port of this interpretation are the observations that (i)
ultrastructually uniform OECs reside throughout the
full depth of the ONL (Au et al., 2002; Valverde et al.,
1992; Doucette, 1989), (ii) in vitro studies have demon-
strated that expression of widely used OEC markers,
e.g. p75NTR , can be regulated by OEC interaction with
extrinsic factors such as axonal contact (Wewetzer et al.,
2005; Ramón-Cueto et al., 1993; Vickland et al., 1991),
and (iii) OECs display morphological plasticity in vitro
(Vincent et al., 2003; Van Den Pol & Santarelli, 2003) and
following transplantation into injured CNS tissue (to be
discussed below).
Relationship between olfactory ensheathing cells
and developing olfactory nerves
The axon pathfinding of olfactory neurons involves a
range of guidance mechanisms which are still the sub-
ject of intense research (reviewed by Key & St John,
2002). In mammals, the direct role of OECs in this pro-
cess is yet to be clearly defined, although glia cells that
play a key role in sorting olfactory axons are present in
invertebrates such as the moth, Manduca sexta (Rossler
et al., 1999). Olfactory axon growth in mammals takes
place in a series of steps, regulated by distinct guid-
ance cues some of which are provided by OECs. First,
axogenesis occurs and new axons emerge from the ol-
factory epithelium into the lamina propria. Axons from
the same neuroepithelial zone fasciculate into olfactory
nerve bundles, which are enveloped by cytoplasmic
processes of OECs along their trajectory in the lamina
propria. As the axons approach the bulb, they segregate
broadly into dorsally and ventrally projecting nerve
Morphological and functional plasticity of olfactory ensheathing cells
bundles. Once within the ONL, the olfactory nerves
defasciculate and sort into smaller bundles. It has been
shown that axons expressing the same odorant receptor
gene converge and target to specific glomeruli which
are topographically fixed (Belluscio et al., 2002; Bozza
et al., 2002; Mombaerts et al., 1996). The elongation of
olfactory axons in the lamina propria may be attributed
largely to the behaviour of OECs (Key & St John, 2002),
whereas OEC involvement in the targeting of axon bun-
dles in the ONL to specific glomeruli is unlikely.
The trajectory of olfactory nerves established during
development are formed by conduits of OEC processes
expressing cell adhesion and extracellular molecules
such as galectin-1, β2-laminin, NCAM, E-NCAM, L1
and collagen IV (Crandall et al., 2000; Miragall et al.,
1989; Whitesides & LaMantia, 1996). These growth-
permissive tracks laid down by OECs are thought to fa-
cilitate large-scale fasciculation and elongation of new
axons towards their central targets (Chuah & Au, 1993;
Key & St John, 2002; Whitesides & LaMantia, 1996).
However, axon elongation is optimised in conditions
which limit adhesion (St John et al., 2000), such as
by the expression of polysialic acid (PSA) on NCAM
(E-NCAM) and EphA5 (St John et al., 2000). These
molecules are expressed during the developmental pe-
riod corresponding to massive axon growth and are
postnatally downregulated (Miragall et al., 1989; St John
et al., 2000). EphA5 is expressed by olfactory neurons
and OECs and is essential for olfactory axon elongation
in vitro (St John et al., 2000). EphA5 belongs to a family
of repulsive factors but is thought in this case to facili-
tate direct and unbranching axon growth by regulating
adhesion strength (St John et al., 2000).
In addition to acting as a suitable substratum, OECs
are thought to migrate in tandem with the growing ol-
factory axons. The guidance mechanisms controlling
OEC migration to the presumptive olfactory bulb dur-
ing development are ultimately responsible for the di-
rectional guidance of the pioneer axons and subsequent
establishment of the olfactory nerve tracts (Key & St
John, 2002). This migration may be mediated by a bulb-
derived chemoattractant (Liu et al., 1995), similar to that
required by olfactory axons in the moth, Manduca sexta
(Oland et al., 2003). The same or related factors may be
responsible in rats for attracting neuronal precursors
from the subventricular zone to their olfactory destina-
tions (Liu & Rao, 2003).
At the distal end of olfactory nerves, OECs create an
axon-sorting domain in the ventral ONL where olfac-
tory axons initially cross the PNS-CNS boundary (Cran-
dall et al., 2000). Olfactory ensheathing cells differen-
tially express galectin-1 and its ligand β2-laminin and
the chemorepulsive factor Sema3A in the ONL (Cran-
dall et al., 2000; Schwarting et al., 2000; Pasterkamp et al.,
1998). The subpopulation of olfactory axons express-
ing the Sema3A receptor, neuropilin-1, is repelled from
the ventral Sema3A-positive region, while the subpop-
71
ulation expressing lactosamine-containing glycans con-
verges on this region which has high expression of all
three molecules (Crandall et al., 2000; Schwarting et al.,
2000). Olfactory ensheathing cells also differentially ex-
press galectin-7 (Storan et al., 2004) and p75NTR (Cran-
dall et al., 2000) during development, although roles
for these putative guidance molecules have not been
established.
The role of OECs in guiding olfactory axonal growth
may be attributed also to their expression of glia-
derived nexin, a protease inhibitor which also possesses
axonal-promoting properties (Reinhard et al., 1988). In-
terestingly, the ONL of the olfactory bulb is the only
site in the mammalian CNS where ACs also express
glia-derived nexin (Scotti et al., 1994). Whether this re-
flects some special property of olfactory bulb ACs as
a result of their close spatial association with OECs, or
whether OEC function is influenced by surrounding
ACs remains to be elucidated.
It can be summarised from these studies that OECs
express a complex repertoire of molecules that are
known to be developmentally significant and assume
functionally crucial roles both while they are a com-
ponent of the peripheral olfactory nerves as well as a
cellular constituent of the ONL in the bulb. In light of
their expression of axon-promoting molecules and the
general guidance role that they play in the development
of the primary olfactory pathway, OECs have become
an obvious experimental tool in the repair of the injured
CNS. The following sections will discuss structural in-
teraction between OECs and other cell types in such a
novel environment.
Olfactory ensheathing cells in injured central
nervous tissue
Several studies have indicated that OEC transplanta-
tion holds promise as a potential therapy for spinal
cord injury, although other studies have suggested
that OECs may not always be beneficial. Compari-
son between studies is made difficult by a multitude
of approaches by the different research groups, for
example the various purification strategies and cri-
teria used for defining OECs (Table 1). In the most
successful experiments, adult rats that received OEC
transplants recovered climbing ability and sensori-
motor reflexes after complete spinal cord transection
(Ramón-Cueto et al., 2000), respiratory function after
cervical hemisection (Li et al., 2003a; Polentes et al.,
2004) and bladder control after lumbosacral dorsal root
transection (Pascual et al., 2002). This functional re-
covery correlated with anatomical and activity-based
regeneration in the associated axon tracts, including
long distance regrowth of some axons (Pascual et al.,
2002; Polentes et al., 2004; Ramón-Cueto et al., 2000;
Ramón-Cueto & Nieto-Sampedro, 1994; Ramón-Cueto
et al., 1998). Furthermore, OECs transplanted from
72
rats, dogs, pigs and humans into demyelinating le-
sions in the rat spinal cord promoted remyelination
of axons and restored impulse conduction (Barnett
et al., 2000; Imaizumi et al., 2000a; Imaizumi et al.,
2000b; Imaizumi et al., 1998; Kato et al., 2000; Li et al.,
1998; Smith et al., 2002). Similar results were obtained
when OECs were transplanted from pig into non-
human primate demyelinating lesions (Radtke et al.,
2004).
Although OECs are able to promote regeneration in
the CNS (Guntinas-Lichius et al., 2002; Li et al., 2003b;
Verdú et al., 1999), the precise mechanisms mediating
this process remain ill-defined. Using an in vitro model
of axonal injury, it has been shown that OECs formed a
physical substrate for the growth of post-injury neurite
sprouts (Chung et al., 2004). Neurite sprouting was most
exuberant when OECs were allowed to contact directly
the injured neurons. If OECs and injured neurons were
physically separated but allowed to share the same cul-
ture medium, neurite sprouting was reduced but not
blocked completely. Hence, the results suggested that
both secretion of soluble factors and direct membrane
interaction with injured axons are likely to be involved
in OEC-induced axon regeneration (Chung et al., 2004).
In a coculture assay for neurite growth of retinal gan-
glion cells, p75NTR -immunoselected OECs were found
to be more efficacious in promoting neurite outgrowth
than p75NTR -negative cells from the same tissue or
than a mixed population (Kumar et al., 2005). Whether
p75NTR -positive cells were harvested from the ONL or
from peripherally located olfactory nerve rootlets made
no difference to neurite outgrowth in this assay (Kumar
et al., 2005). The precise function of p75NTR expressed
by OECs in promoting neurite growth remains to be
elucidated.
In in vivo situations, where OECs necessarily interact
with a greater variety of cell types and within a three
dimensional matrix, additional factors come into play.
It appears that restricted migration and optimal phys-
ical orientation of OECs at the injury site may play a
role in inducing a favourable environment for regener-
ation. (Li et al., 1998). Interestingly, studies that involve
transplantation of OECs into CNS regions other than
the spinal cord, have reported more widespread mi-
gration of OECs (Gudiño-Cabrera & Nieto-Sampedro,
1996; Gudiño-Cabrera et al., 2000). For example, when
OECs were transplanted to the hippocampal CA1 re-
gion, they could be found one month later in distant
loci such as the laterodorsal thalamic nuclei, internal
capsule and arcuate nucleus (Gudiño-Cabrera & Nieto-
Sampedro, 1996). This migratory behaviour was shown
to be detrimental to regeneration when OECs were im-
planted into the brainstem and they migrated in a di-
rection opposite to the intended axonal target (Gudiño-
Cabrera et al., 2000).
Migration of OECs appears to be more restricted in
the spinal cord, particularly in cases of complete tran-
V I N C E N T, W E S T and C H U A H
section (Lee et al., 2004). In studies where axonal re-
generation was successful, OECs adopted a spindle-
shaped morphology forming a bridge or scaffold, facil-
itating the growth of axons across the glial scar (Boruch
et al., 2001; Li et al., 1998; Pérez-Bouza et al., 1998). How-
ever, the efficacy of OECs is not consistently guaranteed
(Gudiño-Cabrera et al., 2000; Takami et al., 2002) and is
highly contentious at the dorsal root entry zone, be-
ing supported by many studies (Li et al., 2004; Navarro
et al., 1999; Pascual et al., 2002; Ramón-Cueto & Nieto-
Sampedro, 1994; Taylor et al., 2001) but disputed by
others (Gomez et al., 2003; Ramer et al., 2004a; Riddell
et al., 2004). These discrepancies warrant further inves-
tigation into the cellular mechanisms of OEC-mediated
repair. It is possible that the degree of inflammation
present in the injury site coupled with the manner of
interaction between OECs and astrocytes influences the
extent of repair. Some researchers have argued that
transplantation of OECs by means of cell injections
(Ramer et al., 2004a; Riddell et al., 2004) causes exten-
sive damage of spinal cord tissue which impedes axonal
repair and interferes with optimal interaction between
OECs and ACs (Li et al., 2004). Instead, the application
of OECs within an endogenous matrix to the cut sur-
face of a dorsal root and subsequent stabilisation with
fibrin glue afforded the optimal condition for repair (Li
et al., 2003a; Li et al., 2004).
Recent in vitro studies have shown that there is a
marked difference in the way that OECs and SCs in-
teract with ACs (Lakatos et al., 2000; Van Den Pol
& Santarelli, 2003). Olfactory ensheathing cells were
shown to migrate and intersperse with ACs, whereas
SCs and ACs remained segregated (Lakatos et al., 2000).
Recent evidence suggests that differential regulation of
N-cadherin in OECs and SCs may be the reason for
their differing interaction with ACs (Fairless et al., 2005).
The affinity which OECs show towards ACs could un-
derly their ability to induce changes in host reactive
ACs at the injury site. It has been shown that trans-
planted OECs are capable of migrating both longitudi-
nally and laterally within the injured CNS and can in-
termingle with ACs within the glial scar (Ramón-Cueto
et al., 1998). Furthermore, their ability to migrate within
the injured spinal cord is greater than that of SCs (Li
et al., 1998). However, it should be noted that migra-
tion of OECs appears to be limited by the extent of the
glial scar. In vivo tracking of OECs labelled with super-
paramagnetic iron oxide nanoparticles using magnetic
resonance imaging showed that OECs were not able
to cross the glial scar present at the site of complete
spinal transection (Lee et al., 2004). Nevertheless, there
is general agreement that compared to those which
received SCs, injured spinal cords which were trans-
planted with OECs had comparatively less astrogliosis
as measured by hypertrophy and expression of chon-
droitin sulfate proteoglycan and GFAP (Garcia-Alias
et al., 2004; Lakatos et al., 2003).
Morphological and functional plasticity of olfactory ensheathing cells 73

Fig. 2. Scanning electron micrograph of underside of porous inserts facing astrocytes (A). The pores measure 1 µm in diameter
and two of the pores demonstrate the presence of OEC processes emerging from the other side. In the rare instance, entire OECs
had squeezed themselves through the pores and are apparently hanging on by their narrow processes (B). Scale bar = 1 µm
(A) and 4 µm (B).

In a study involving lesion to the dorsal root en-
try zone in which there was successful regeneration of
about 10% of axotomised fibres, it was shown that trans-
planted OECs interacted with host ACs and SCs of the
dorsal root to form a ladder-like structure that allowed
ingrowth of nerve fibres across the normally inhibitory
PNS-CNS interface (Li et al., 2004). Using an in vitro
model which allows populations of cells to be physi-
cally separated but sharing the same culture medium
(Chung et al., 2004), we have obtained some preliminary
results that suggest that interaction between OECs and
reactive astrocytes may be mediated by soluble factors.
OECs were cultured from neonatal rat ONL and ol-
factory mucosa and purified as previously described.
(Vincent et al., 2003), and with approval from the Ani-
mal Experimentation Ethics Committee of the Univer-
sity of Tasmania. Olfactory ensheathing cells were cul-
tured overnight in porous Falcon Cell Culture Inserts
(catalog # 353104, Becton Dickinson Labware, Franklin
Lakes, NJ) overlying ACs, which were pre-activated
by scratch wounding and exposure to 10 ng/mL trans-
forming growth factor β (O’Toole et al., 2005). The
porous inserts allowed the two cell types to share the
same medium in the absence of direct contact. Follow-
ing 24 hours’ culture, processes of some OECs could be
observed traversing the 1 µm diameter pores and ex-
tending towards the ACs (Fig. 2A). In rare instances
entire OECs were able to squeeze through the 1 µm di-
ameter pores and to hang on to the underside of the
inserts by their processes (Fig. 2B).
In summary, transplanted OECs display morpholog-
ical plasticity in response to as yet unidentified cues
from the novel injured CNS environment. It is likely that
the morphological plasticity demonstrated by OECs re-
flects an equally robust functional plasticity in their in-
teraction with CNS tissues.
Other possible functional roles of olfactory
ensheathing cells
It is well accepted that olfactory neurons lining the
nasal cavity are continuously renewed throughout an
74 V I N C E N T, W E S T and C H U A H

animal’s life (Graziadei & Graziadei, 1979); some of vent the turbidity induced in the culture medium when
them have a lifespan of about one month while oth- the latter is infected with attenuated Staphylococcus au-
ers are known to survive for much longer (Mackay-Sim reus and Escherichia coli (JM109 strain) (Vincent et al.,
& Kittel, 1990). As mature olfactory neurons die, they 2005). Further investigation was done to test whether
are replaced by a new population of cells derived from OECs were capable of phagocytosing whole bacteria
globose basal cells residing in the olfactory epithelium in vitro. Fluorescently-labelled Microccoccus luteus (M.
(Mackay-Sim & Kittel, 1991). In the unperturbed ani- luteus; dead, Gram-positive; EnzCheck Lysozyme As-
mal, it has been observed by electron microscopy that say kit, Molecular Probes, Eugene, OR) were incubated
small numbers of macrophages are present in the olfac- with OECs for 24 h and the living cultures were viewed
tory epithelium. (Graziadei & Graziadei, 1979). These under fluorescence microscopy. M. luteus were associ-
macrophages were assumed to be responsible for dis- ated preferentially with the cell bodies of many OECs
posing of degenerating epithelial cells under normal in each culture, particularly surrounding the nucleus
physiological conditions. However, they are not the (Fig. 3A and B). Interestingly, internalised M. luteus ap-
only cell type that is capable of phagocytosing cel- peared to have been protected from cross-reaction with
lular debris in the olfactory system. Bulbectomy and
infection by neurotropic pathogens induce rapid and
massive death of olfactory neurons. Removal of the re-
sulting cell debris and pathogens in the acute phase is
marked by an increase in the number of macrophages
infiltrating the degenerating epithelium. (Suzuki et al.,
1995). In addition, supporting cells of the olfactory ep-
ithelium also become phagocytic as indicated by the
presence of phagosomes and apoptotic bodies in the
cytoplasm (Suzuki et al., 1996). Not only are they ac-
tive during the acute phase of degeneration, phagocytic
supporting cells can be observed up to two months after
bulbectomy (Suzuki et al., 1996).
Whilst macrophages and supporting cells are in-
volved in clearance of dead cell bodies in the epithe-
lium, OECs appear to contribute to phagocytosis of ax-
onal debris in the lamina propria and olfactory bulb
(Chuah et al., 1995). Electron microscopy showed that
following zinc sulphate irrigation of the nasal cav-
ity which destroyed much of the olfactory epithelium,
some OECs forming the glia limitans of the olfactory
bulb were observed to contain phagosomes and cellu-
lar debris in their cytoplasm (Chuah et al., 1995). The
phagocytic activity of bulb-derived OECs has been re-
cently confirmed in an in vitro study which showed
that the O4 immunoreactivity of OECs was actually
due to O4-positive axonal fragments adhering to OECs
(Wewetzer et al., 2005). Using a novel two-step label-
ing protocol based on antibody internalisation, Wewet-
zer and co-workers showed that the O4-positive ax-
onal fragments were phagocytosed by OECs, and as
immunoreactivity for O4 decreased, the OECs upregu-
lated their expression of p75NTR .
Based on these findings, it is logical to ask if OECs
are capable of showing anti-bacterial activity, such as
when infected olfactory axons degenerate and release Fig. 3. Green fluorescently labeled M.luteus surround the nu-
pathogens, thus acting as a member of the innate im- cleus of OECs stained by Hoescht Blue dye (A); cytoplasm
mune system. Given that OECs are present in the lam- of the OECs stain positive for S100 (B). In (C) immunostain-
ina propria beneath the olfactory epithelium, it would ing for Lyz show different intensities of labeling in two OECs.
be reasonable to assume that they probably form a The M.luteus adhering on the external surface of the unperme-
line of defence secondary to olfactory supporting cells abilised OECs or to the coverslip show cross-reactivity with
and macrophages. In preliminary experiments from our anti-Lyz (bright orange) while the internalized M.luteus are
laboratory, it was found that OECs were able to pre- fluorescently green. Scale bar = 10 µm.
Morphological and functional plasticity of olfactory ensheathing cells
anti-lysozyme (Lyz) antibodies in the unpermeabilised
cells (Fig. 3C), in contrast to M. luteus found attached
to the external surface of OECs or to the coverslip.
The finding that OECs could have antibacterial ac-
tivity in culture is intriguing given that the olfactory
system is the only neuroepithelium in the body that is
directly exposed to the external environment. Mecha-
nisms which are known to protect this tissue from in-
fection include immunogenic mucosal secretion, and
lactoferrin and Lyz secretion from Bowman’s glands
(Getchell & Getchell, 1991). In addition, recent studies
from our laboratory have shown that OECs produce
mRNA for Lyz and are immunopositive for Lyz protein
both in vivo and in vitro (Vincent et al., 2005). Although
it was not stated by the authors, a previous immuno-
histochemical study in the rat and salamander showed
that OECs appeared to be immunopositive for Lyz (see
Fig. 6h in Getchell & Getchell, 1991). A role for Lyz
activity in OECs is consistent with the susceptibility
of the primary olfactory pathway to external chemi-
cal and pathogenic insults. A recent study showed that
pneumococcal infection can occur from the olfactory
epithelium to the olfactory bulb via transport along ol-
factory nerves, but that this route is limited by an un-
known mechanism (van Ginkel et al., 2003). If OECs
can be proved to contribute to a neuroprotective role in
the olfactory system, in the future it may be possible to
prime or activate them by intranasal delivery of specific
drugs or genes (Frey, 2002; Thorne et al., 2004) in order to
protect people at high risk of contracting meningitis or
encephalitis.
In a recent microarray study, factors involved in the
immune response and inflammation have been de-
tected as transcripts enriched in OECs, including the
chemokines Cxcl1 (also known as Gro1) and Ccl2 (also
known as MCP1). Ccl2 is a monocyte chemoattractant
and Cxcl1 is a neutrophil chemoattractant that is also in-
volved in glial development (Dong & Benveniste, 2001).
The protein levels for these factors are low or absent in
OECs in vitro and in normal olfactory tissue (Vincent
et al., 2005), but this is consistent with the expression
of Ccl2 in Schwann cells, which is normally low in the
sciatic nerve and is upregulated in response to injury
(Toews et al., 1998). Astrocytes in the spinal cord pro-
duce Cxcl1 in a spatiotemporal manner to help direct
oligodendrocyte precursor maturation and migration
(Tran & Miller, 2003). Although immunohistochemical
staining showed that OECs do not appear to produce
detectable amounts of Ccl2 or Cxcl1 proteins in the nor-
mal adult olfactory system, a role for these factors may
exist when these cells are transplanted into the injury
site in the CNS.
Furthermore, it is likely that OECs are themselves
capable of responding to immune factors, as suggested
by their high level of expression for the interleukin 6
(IL-6)-responsive transcription factor, Cebpb (Vincent
et al., 2005). This notion is supported by a study which
75
reported that in the aftermath of bulbectomy and fol-
lowing infiltration of IL-6 producing macrophages into
the olfactory lamina propria, OECs were shown to up-
regulate their expression of IL-6 receptors (Nan et al.,
2001). These authors proposed that IL-6 may be in-
volved in initiating intracellular signalling processes
that ultimately lead to OECs establishing extracellu-
lar matrices which facilitate olfactory axonal regrowth
(Nan et al., 2001). Such a capability could be function-
ally significant as well, when OECs are transplanted
into an acute injury site in the CNS.
Another protein, which could prove to be function-
ally significant in the context of OECs as a form of cell
therapy in CNS injury, is connective tissue growth fac-
tor (Ctgf). In the unperturbed olfactory system, Ctgf has
been observed in the embryonic mouse olfactory ep-
ithelium (Surveyor & Brigstock, 1999). Ctgf is involved
in diverse processes in the nervous system which in-
clude angiogenesis, fibrotic and glial scarring, and ex-
tracellular matrix remodelling in normal and injured
tissue. (Hertel et al., 2000; Schwab et al., 2001). In a re-
cent study, it was reported that the region of the dorsal
column in which OECs were injected had deposits of
laminin and showed the presence of numerous new
blood vessels. (Ramer et al., 2004b). It is tempting to
speculate that proteins expressed by OECs, such as Ctgf
and vascular endothelial growth factor (Au & Roskams,
2003), may be involved in the reported neoangiogene-
sis. Intriguingly, although neovascularisation is gener-
ally thought to be beneficial to repair, in this particu-
lar study, axotomised dorsal roots failed to regenerate
(Ramer et al., 2004b).
An immune function in OECs may have significant
implications for the transplantation of these cells as a
therapy for CNS injury. Physical trauma of the CNS
is often accompanied by a massive inflammatory re-
sponse which is thought to cause secondary damage
leading to neuron loss (Bareyre & Schwab, 2003; Blight,
1992; Carmel et al., 2001). The anti-inflammatory drug
methylprednisolone is used in the treatment of hu-
man spinal cord injury in an attempt to manage this
secondary damage. Concievably, transplanted OECs
may promote regeneration via a similar mechanism,
by modulating neuroinflammation in the injured CNS.
Indeed, OECs used in a combination treatment with
methylprednisolone in lesioned rat spinal cord were
found to have a synergistic effect on axonal regenera-
tion and functional recovery (Nash et al., 2002).
In summary, increasing understanding of the func-
tional plasticity of OECs may well reveal that this
unique glial cell could have multiple functions extend-
ing beyond its role in the normal olfactory system.
There is now much experimental evidence that shows
the regenerative property of the olfactory system can
replace the inhibitory nature of the injured CNS by
OEC transplantation. Whilst their use in promoting re-
pair in experimentally induced CNS injury reflects their
76
versatility, their future development into a reliable clin-
ical therapy can only be assured by a better understand-
ing of their cell biology.
Acknowledgments
Research conducted by the authors was supported by
grants from the National Health and Medical Research
Council and the International Institute for Research in
Paraplegia. MIC would like to express her gratitude to
Albert Farbman for his mentoring which started her on
a career in basic research.
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