Primary Cell Culture Essay

Title: Primary cell culture of a chick embryo
Aim: To establish and maintain the primary cell culture of a chick embryo
Theory: Historically, early attempts at tissue culturing used biological culture medium such as blood serum, clotted lymph or tissue extract to grow cells outside the body. Tissue culturing is a biological research that involves the fragmenting of tissue from an animal or plant. These tissues are transferred to an artificial environment to test their ability to survive and function. The artificial environment or medium must contain proper proportions of the necessary nutrients, amino acids, and a balanced pH for the cells to be studied. Cultures are can grown either as single layers of cells on a glass or plastic surface or as a suspension in

20 ml DBSS was pipetted out and added into the dish.
8. The embryo was dissected using two scalpels and the organs arranged around the periphery of the dish.
9. 1ml (1000 µl) of 0.25% Trypsin was pipetted out and added to a clean test tube. A piece of the dissected tissue was transferred into the test tube containing trypsin using forceps and the tube closed.
10. The tissue was incubated at 40C for 6- 18 hours in a refrigerator.
11. After incubation, the trypsin from the test tubes was carefully removed without disturbing the tissue and further incubated in the residual trypsin for 15–20 min at 370C.
12. 2ml of media was added to two 60mm petriplate and to the test tube containing tissue and the residual trypsin. The media was then pipetted up and down in the test tube to disperse the tissues.
13. The tissue was equally transferred into petriplates using pipettes. The media was then mixed in the Petri plate using a micro pipette.
14. The cells were incubated in the CO2 incubator for 12 hours.
15. The cells were viewed through the inverted microscope after an overnight incubation.
16. Confluency of plate was checked, and the cells again incubated in the CO2 incubator for 5-7 days to get a confluent