Hela Cell Culture Media Protocol

Replace this immediately by carefully pouring an equal volume of pre warmed fresh culture media into the flask.

Hela cell culture media protocol.

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 cell culture protocol hela cells can be grown to the appropriate density usually 70 in a humidified chamber at 37 c 5 co2. Aspirate medium from the flask. For example put the required volumes of cell media into.

Before starting the protocol. Prepare the new dishes and or six well plates which will be used for the new split. Cell culture of 7721 or hela cells protocol method by jiao wang. The cell line was found to be remarkably durable and.

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. Splitting hela cells the following protocol is for passaging hela cells that are 70 confluent in a 10 cm dish. 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. However depending on each cell type above mentioned reagent condition changes.

Bring cell media pbs and trypsin to room temperature. As of now i know to culture any cells we need media antibiotics and many other reagents. Incubate cells at 37 c with 5 co2. 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.

Cells are maintained in 10cm dishes at 37 c and 4 co2until 60 80 confuency. Using cell scraper gently scrape the cells off the bottom of the flask into the media. Renew growth medium 2 3 times a week. Remove and discard culture medium.

Hela are very tough and will survive a. Hela cell doubling time is 24 hours. Subculture protocol for hela cells. Hela ˈ h iː l ɑː.

It is the oldest and most commonly used human cell line. Add 10 ml sterile 1x pbs and rinse cells by pipette. Hela cells are adherent cells grown in eagle s mem emem modified with 10 fbs. 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.

Also hela or hela is an immortal cell line used in scientific research.