I have trouble growing a cell line that I had just purchased from a cell line distributor. The cells I bought were produced in 2016 and when they were seedling they couldn't grow. There were some (not more than 10 single cells) that can grow larger in size but cannot divide and eventually die. Does anyone have experience in overcoming problems like mine? Are there any compounds that can increase the viability and stimulate cell viability and cell division?
In general, when mammalian cells don't grow "fast enough" you can help them by adding extra serum and non-essential amino acids, but probably someone who has worked with a similar cell line knows best. Could you give more information about the cell line (name, cell type) and provider?
Ten individual cells after thawing are too few, and such selection might affect your conclusions and reproducibility out of your lab.
If freezing and thawing was performed appropriately, cells should grow in the suggested culture conditions (i.e. medium, additives, 37ºC, CO2, etc), and if not, you should apply guaranty.
Dear Harto!
Of course there are such connections. For example, growth factors - epidermal growth factor (EGF), fibroblast growth factor-2 (basic) - FGF2, many interleukins, such as: IL2, IL4, etc.
You need the name of your line and look for protocols for culturing these cells. Because the proliferation of a certain line requires its own growth factors
Hello Harto
You have not mention the cell line that is causing a problem. Nevertheless, l am making a mention of a few compounds that could help you solve your problem.
You can use interleukin-2 which is a pleiotropic cytokine that helps in T-cell growth.
Phorbol Myristate Acetate (PMA) and ionomycin help to induce T cell proliferation.
Granulocyte Colony Stimulating Factor (GCSF) is a pro-inflammatory cytokine that stimulates myeloid stem cell proliferation. It can also act as a growth factor to certain extent.
Then there are other growth factors like Epidermal Growth Factor (EGF) and Platelet Derived Growth Factor (PDGF) that can help in the proliferation of a number of different cell types.
I hope this information is helpful.
Regards.
Thank you very much Mr. Luis Gómez-Morales; Alexandr Chernov ;
The cell line is Brin-BD (a pancreatic cell line)
We used to perform appropriately freezing and thawing since that works are routine in our lab. We have already complied the included protocol: the cells were grown in the suggested culture conditions (RPMI medium and we also have tried with DMEM, 37ºC, CO2, etc) but we have a very poor result (fail).
Thank you very much, Mr. Malcolm Nobre for your valuable and helpful answer
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