Splitting hela cells the following protocol is for passaging hela cells that are 70 confluent in a 10 cm dish.
Hela cell culture media protocol.
Bring cell media pbs and trypsin to room temperature.
However depending on each cell type above mentioned reagent condition changes.
It is the oldest and most commonly used human cell line.
Before starting the protocol.
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.
Remove and discard culture medium.
Cell culture of 7721 or hela cells protocol method by jiao wang.
Check all the cells have come off by inspecting the base of the flask before moving on.
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.
Prepare the new dishes and or six well plates which will be used for the new split.
Hela cell doubling time is 24 hours.
Add 10 ml sterile 1x pbs and rinse cells by pipette.
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 cells are adherent cells grown in eagle s mem emem modified with 10 fbs.
Cells are maintained in 10cm dishes at 37 c and 4 co2until 60 80 confuency.
Aspirate medium from the flask.
Subculture protocol for hela cells.
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.
Also hela or hela is an immortal cell line used in scientific research.
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.
Renew growth medium 2 3 times a week.
You can wean them onto other media at a later time if you like.
Replace this immediately by carefully pouring an equal volume of pre warmed fresh culture media into the flask.
Hela are very tough and will survive a.
Using cell scraper gently scrape the cells off the bottom of the flask into the media.
Hela cell culture protocol hela cells can be grown to the appropriate density usually 70 in a humidified chamber at 37 c 5 co2.
Hela ˈ h iː l ɑː.
Hela cells in many labs are cultured in 75 cm2 flasks or medium flasks.
