Please, I need some instruction about the cell line culture; I work on SH-Y-5Y, and HEK293, using different concentrations of chemotherapy to achieve mutation.
Hi, everyone.
Please, I need some instruction about the cell line culture; I work on SH-Y-5Y, and HEK293, using different concentrations of chemotherapy to achieve mutation.
First of all, What is the cell line protocol for (SH-Y-5Y), &(HEK23) ?
2nd, how can I choice the concentration variation, for example, is that depends on the IC50, and what is the best way to know the IC50 for cisplatin for example?
What is the protocol to deals with these kinds of cell lines?
I will high appreciate answers and advices with an article, book, or section.
Please clarify the cell line you are trying to grow, SH-Y-5Y or SH-SY5Y? Media for HEK293 is Eagle's Minimum Essential Medium with 10% FBS by ATCC. Media for SH-SY5Y is Eagle's Minimum Essential Medium/F12 or DMEM/F12 with 10% FBS. Always add glutamine for your cells.
IC50 for commonly used drugs are available by Google search (1st link attached is IC50 for cisplatin). To test IC50 of a drug (2nd link), you need to do a serial dilution of the drug using cell proliferation assay such as using MTT or ELISA in a 96 well format. First, do a Google search to see if others have tested the drug of interest to get an idea what is the range of concentration used and then apply to your cell line. If you can't find any references, try 1 nM, 10 nM, 100 nM, 1 uM, 10 uM, 100 uM first. If you don't see any effect, it is not a good drug to spend your time, pick another one to test. Hope these helped. Good luck!
http://www.jbc.org/content/suppl/2009/06/26/M109.026583.DC1/jbc.M109.026583-2.pdf
http://www.sciencegateway.org/protocols/cellbio/drug/hcic50.htm
Dear Ying Henderson;
I am highly appreciated your time and your help, I am working on SH-SY5Y & HEK 293 to identify the mutation(s) gene(s) when I treat these cell line with a chemotherapy, it is a big job ,unfortunately just fewer people in my university ,providing some support to this project, my aim is a novel idea in cancer research I could not discuss it in this small size area. Best regards and thanks again.
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