I have to use pre-coated flasks for some of my cell lines (BEAS-2B, THLE-2). Can anybody help me with the flask coating (BSA, collagen and fibronectin)? For how long can I store the prepared coating solution? How long should I leave the coating solution in the flask? How long can I store the coated flasks before I begin the cell culture?
Here is the protocol for coating the flasks with 0.1 % gelation
1. Weight 1 gm gelatin, put into 1 liter of deionized water and autoclave.
2. Pour 5-10 ml of autoclaved gelatin solution into 10 ml pertiplate/75mm mammalian tissue culture grade plate/ flask (in laminar flow Hood)
3. Cover the entire bottom surface of the plate/flasks with gelatin solution.
4. Put the flask/plate in 37 C CO2 incubator for half and hour.
5. Remove the flask from the incubator, take it to Laminar Flow Hood, remove gelatin solution, wash/rinse with PBS/HBSS and directly use it for cell culture without drying.
6. After washing with PBS, you can store the plate/flasks in refrigerator (4oC) for upto one week before use without drying the plate.
7. Once again the plate must not be dried while storing in refrigerator. So, do not remove PBS if you do not want to use the coated plate immediately.
What are the steps for flask coating for THLE- 2 cells . can collagen alone work ..?? kindly help me out
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