Cell line characterisation
- 1. CELL LINE CHARACTERISATION Presentation by – Teena Roll no – 5 Submitted to –kurukshetra department DR. Sulekha Chahal .
- 2. Need for characterization • Demonstration of the absence of cross contamination • Confirmation of species of origin • Determination of whether the cell line is transformed or not. • Indication to whether cell line is prone to genetic instability and phenotypic variations. • Identification of specific cell line within a group from the same origin.
- 3. AUTHENTICATION • For checking the cross contamination it is necessary to check the authenticity of cell line It can be done in various ways. Some of the methods in general use for cell line characterization are
- 4. Criteria Method Karyotype Chromosome spread with banding Isoenzyme analysis Agar gel electrophoresis Cell surface antigen Immunohistochemistry Cytoskeleton Immunocytochemistry with antibodies to specific cytokeratins DNA Fingerprint Restriction enzyme digest; PAGE ; Satellite DNA probes
- 5. Cell Morphology •Simplest and most directed technique use to identify cells •Shortcomings- cellular morphology varies in response to culture conditions •Comparative observation should always be made at some stage of growth and cell density in same medium and on same substrate.
- 6. Cell morphology study 1)Microscopy It could be use to study the morphology and anatomy of cells 2)Photomicroscopy ie photograph plus microscopy 3)Staining
- 7. Chromosome content •For identifying cell line and relating them species and sex from which they have derived •Also diffrentiate normal and transformed cells •Includes 1. Chromosome banding(casperson etal in 1970) • C banding-stain centromere • Q banding- stain AT rich region • G banding-stain sulphur rich protein part R banding-stain sulphur deficient region 2. Chromosome analysis 3.Chromosome painting