We ordered HSC-F cell line (H.Saimiri immortalised T-cell line from foetal Cynomolgus monkey) from NIBSC (Confidence in Biological Medicines) two years ago. According to centre for AIDS reagents datasheet of HSC-F cell line, culture medium should contain 85% RPMI 1640, 15% Foetal calf serum, IL-2 (10IU) and 50μM β-Mercaptoethanol. I used these ratio of contents for medium in 60 mm plate. But Growth of HSC-F cells occurred slowly such that during one year had not passaged them. Consequently, They started to contaminate and their growth stopped completely. We decided to order again from the same place. Unfortunately I encountered this problem again. I have been trying to maintain and transfect them for seven months. But it is pointless.
Hi Aysu hanim,
As a graduate of METU University (I did my MSc in Biotech there), I am familiar with Turkish language and a strong fan of Turkish food.
I have to ask you please take your health serious when working with chemicals such as:
β-Mercaptoethanol.
Regarding your question:
Manufacturer (NIBSC) recommends that these cells are
initially established a 25cm2 tc flask in approximately 3ml
of media. Growth is sometimes improved by placing the
flask at an angle of 20-30 degrees so that the cells are
concentrated in the corner of the flask. Media should be
added slowly over subsequent days. Only split the cells
one they are quite thick and the media is yellow. Split one
T25 flask into two flasks , and add a small amount of fresh
media. One T25 flask containing a white film of cells (in
approximately 10 mls media) can be frozen into two vials
of cells.
Please see the attached PDF file and let me know if you have further questions or you want me contact the manufacturer/ supplier.
Iyi gunler va calismalarinda basarilar dilerim
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