I need to set CLL proliferation assay, but my CLL cells are not proliferating. I am incubating for 72-96 h in IL4 & bME containing RPMI (reduced serum).
If anybody knows the trick to grow CLLs, pls let me know
Hi Bhawana,
First, are the CLL cells primary cells from patients, or a cell line? If the cells are primary cells, it is quite difficult to proliferate in vitro. If they are from a cell line, why did you use reduced serum? You should use at least 10% of FBS.
Second, have you checked the cell state after thawing? Did they look healthy or unhealthy?
Third, contamination is another common reason for not growing.
Could you see this thesis
The role of toll-like receptor 9 on B-cell activating factor expression and function in normal human B-cells
EYA Abu-rish - 2013 - lra.le.ac.uk
https://lra.le.ac.uk/handle/2381/27908
Best Wishes,
Hi Zhang, These CLLs are isolated from patients, frozen and thawed. At the time of thawing, cell health was good, just i am not able to proliferate them
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