Cell Fusion and

Hybridization
Reporter:
Mapaa, Carrelle Joy T.
Mamontayao, Jessievel
Masugod, Aiza
 Cell Fusion
Fusion of two different cells into a single unit
which contains genes from both the original cell.

The commonly used fusion agents for mapping of

chromosome are Sendai virus, lysolecithin ,liposomes
and polyethylene glycol (PEG).
• The most widely used fusion agent is
Sendai virus, which is inactivated by UV
light or B-propiolactone.
 Selection of Hybrids
 To isolate pure population of Human-
Mouse Heterokaryon a selection procedure
is used that kills both the parental cells
and the homokaryon but allows the human –
mouse hybrid cells to survive and grow.
 In the medium contains a drug aminopterin
, which blocks de novo purine and
pyrimidine biosynthetic pathway of cells.
 The presence of hypoxanthine and
thymidine the cells can overcome the
block by synthezing and pyrimidines using
salvage pathways.
 For cells to grow in HAT medium, two
enzymes, hypoxanthine guanine
phosphoribosyl transferase (HGPRT) and
thymidine kinase (TK) must be functional.
Heterokaryons , has normal HGRPT gene
derived from the mouse genome and TK
from gene from the human genome.
Therefore, only hybrid cell grows.
Chromosome Loss in Hybrid Cells
 When hybrid are formed between rodent
and human cells, human chromosomes are
lost preferentially.
 Loss of chromosome in interspecific
hybrids was noted in fusion between mouse
and rat cells , where chromosomes of rat
were preferentially lost.
 In rodent and human cell fusions, up to
95% of the human chromosomes can be
lost by the time the hybrids are initially
isolated and their chromosome analyzed.
 Stages of Cell Fusion:
Stage I: Specification of the fusion fate
Stage II: Cell attraction attachment and recognition
Stage 3: Execution of Cell Membrane and
Cytoplasmic Fusion
 Types of Cell Fusion
 Non Self Fusion – a cell fusion which allows
the segregation of genes in new combinations
into a single cell.
 Self-Fusion –
 Auto-Fusion
 Somatic Hybridization
• Development of hybrid plants through the
fusion of somatic protoplasts of two different
plant species/varieties.
Somatic Hybridization Technique
• Isolation of Protoplasts
• Fusion of the Protoplasts of desired
species/varieties
• Identification and Selection of somatic
hybrid cells.
• Culture of the hybrid cells.
• Regeneration of Hybrid Plants
 Application of Somatic Hybridization
• Opened new possibilities for the in vitro
genetic manipulation of plants to improve the
crops.
• Helped in the successful transfer of some
disease resistance genes.
• Environmental Tolerance
• Quality Character
 Problems and Limitations
• Fertile and visible seeds are not always
produced by plants .
• The production of viable somatic hybrids is
not possible in all instances even though
Protoplasts fusion between different species
or genus is easy.