1. CEAC514 Assoc. Prof. Dr. Yasemin G. İşgör
Tissue Cell Culture Technology: Seeding, passaging, harvesting and storing (cryopreservation) of cells
CEAC514
Assoc. Prof. Dr. Yasemin G. İşgör

2. VARIOUS APPLICATIONS OF TISSUE CULTURE TECHNIQUES

3. CELL CULTURE COLLECTIONS (available sources)
General culture parameters-2
CELL CULTURE COLLECTIONS (available sources) .

4. General methods and culture parameters-2
Primary culture
Cells when surgically or enzymatically removed from an organism and placed in suitable culture environment will attach and grow are called as primary culture where the Primary cells have a finite life span.
Primary culture contains a very heterogeneous population of cells (different types of cells present)
Cells such as macrophages and neurons do not divide in vitro so can be used as primary cultures

5. Primary culture (example)
Stage of culture after the cells are isolated from the tissue and proliferated under the appropriate conditions until they completely occupy the substrate
Monolayer – Reach Confluence
Then need to be subcultured (Passaged)
Split & Move to fresh medium, new vessels
Chinese Hamster Ovary Cells (CHO)

6. Preparıng primary culture from mınced tıssue (example)

7. Primary culture: basıc Method
Chop the tissue in media
Digest w collagenase /
Wash with buffer and centrifuge, then
discard supernatant
Use trypsin to
Separate primary
cells
Seed cells at low density
Harvest cells before confluence

8. ISOLATION OF CELLS FOR CULTURING

9. MONOLAYER PROPOGATION

10. Thawing frozen cell stock (general protocol)

11. SUBculturING
Harvesting cells with trypsın-edta
FREEZING STOCKS

12. storing (cryopreservation) of cells
Liquid Nitrogen Cryogen:
The metal tank suitable to hold cells at the vapor
Phase of liquid nitrogen