Hela Cell Culture Media Protocol

Check all the cells have come off by inspecting the base of the flask before moving on.

Hela cell culture media protocol. Subculture protocol for hela cells. Add 2 0 to 3 0 ml of trypsin edta solution to flask and observe cells under an inverted microscope until cell layer is dispersed usually within 5 to 15 minutes. Hela are very tough and will survive a. Using cell scraper gently scrape the cells off the bottom of the flask into the media.

For example put the required volumes of cell media into. Hela cells in many labs are cultured in 75 cm2 flasks or medium flasks. Replace this immediately by carefully pouring an equal volume of pre warmed fresh culture media into the flask. Splitting hela cells the following protocol is for passaging hela cells that are 70 confluent in a 10 cm dish.

Incubate cells at 37 c with 5 co2. I am very new to culture of mammalian cells and i have to design a protocol as a part of my lab work can any one explain me how do we start a culture of hela cells from frozen cells which would be the best culture medium for hela cells. Bring cell media pbs and trypsin to room temperature. Hela cell culture protocol hela cells can be grown to the appropriate density usually 70 in a humidified chamber at 37 c 5 co2.

Before starting the protocol. Hela cell doubling time is 24 hours. Hela cells are adherent cells grown in eagle s mem emem modified with 10 fbs. Prepare the new dishes and or six well plates which will be used for the new split.

Hela ˈ h iː l ɑː. The cell line was found to be remarkably durable and. 3 0 hela cell culture tet on hela cells are cultured using dmem media gibco supplemented with 10 tet system approved fbs clonothec and 1x pen strep 100 units of penicillin 100 µg of streptomycin. Remove and discard culture medium.

Aspirate medium from the flask. As of now i know to culture any cells we need media antibiotics and many other reagents. Briefly rinse the cell layer with 0 25 w v trypsin 0 53 mm edta solution to remove all traces of serum which contains trypsin inhibitor. Cell culture of 7721 or hela cells protocol method by jiao wang.

Cells are maintained in 10cm dishes at 37 c and 4 co2until 60 80 confuency. It is the oldest and most commonly used human cell line. Renew growth medium 2 3 times a week. The line is derived from cervical cancer cells taken on february 8 1951 from henrietta lacks a 31 year old african american mother of five who died of cancer on october 4 1951.

Add 10 ml sterile 1x pbs and rinse cells by pipette.