Hydrogels

Cells in their physiological setting typically interact with extracellular factors in

three dimensions (3D). The extracellular matrix (ECM) is a complex formulation of
biological polymers composed of peptides, polysaccharides, proteins, enzymes and
signaling molecules that reside outside of cells. Interactions between cells and their
surrounding ECM provide cues that can influence cellular polarization, shape,
motility, differentiation and many other phenotypes. Traditional cell culture is done
on a two-dimensional (2D) substrate, usually polystyrene or glass, and cells grown in
this manner often do not fully recapitulate many phenotypes observed by those
same cell types in vivo. One approach to making cells cultured in vitro more closely
resemble their corresponding physiological tissue type is to embed and culture cells
within a 3D hydrogel that mimics the physiological ECM. This strategy is a subset to
the larger approach of culturing cells in three-dimensions, typically referred to as
3D cell culture.

Figure 1. Hydrogels enable more biologically-relevant 3D cell culture.

3D HYDROGEL APPLICATIONS
Cells grown in 3D hydrogels have proven valuable for numerous applications.
Traditional immortalized cell lines can be cultured embedded within hydrogels to
form multicellular complexes called spheroids. Spheroids have been demonstrated
to provide a wealth of information not apparent from the same cells cultured under
2D conditions. Stem cells grown in 3D hydrogels can differentiate into multiple cell
types that closely resemble native tissues. They can be programmed to form 3D
organoids, differentiated mini-organ structures that have been used as highly
predictive, reproducible, and scalable in vitro models of complex physiology. Other
applications of culturing cells within hydrogels have focused on interrogating the
physical properties of the hydrogel to mechano-sensitive behaviors of cells
including the formation of morphological features and cell motility.

NATURALLY-DERIVED 3D HYDROGELS
The most conventional method for mimicking the physiological extracellular matrix
in vitro is to use an extract of native ECM from a biological source. Traditionally, the
ECM extract from Engelbreth-Holm-Swarm (EHS) murine sarcoma basement
membrane has been used for 3D cell culture applications. EHS ECM extract is
conveniently liquid at 4 °C and will gelate at room temperature, allowing cells to be
easily embedded. This product is sold as ECM Gel hydrogel. For some applications,
an ECM extract from human cells is important. MaxGel™ ECM is a human in vitro
derived basement membrane extract (BME).

SYNTHETIC 3D HYDROGELS
Reconstituting a synthetic hydrogel may be the optimal choice for some
applications. A chemically defined hydrogel formulation enhances reproducibility,
and the presence and concentration of media components such as growth factors is
known. In addition, key physical parameters of a hydrogel may be important for
specific applications. These include:

• elastic modulus
• pore size
• hydrogel swelling
• crosslinking
• dissociation conditions

We offer several reagents, kits and protocols to be used for developing synthetic
hydrogels including HyStem® hydrogel scaffolds, TrueGel3D™ hydrogel kits and
Cell-In-a-Box® technology.
Figure 2. Synthetic Extracellular Matrix (ECM)