Hi,
I want to check the effect of bone marrow derived mesenchymal stem cells on the proliferation of leukemic blasts as well as standard cell lines in the presence and absence of standard chemotherapeutic drugs. I havent performed co-culture assays before and want to standardize the experiments. What should i begin with? Should i calculate the IC50 values of BM-MSC and leukemic cells separately? How should i decide the correct proportion of cells to be taken in the co-culture?
Hi Manasi Nagare ,
For co-culturing, you will need to have the same growth media. Try to accommodate the cells on the same growth media. In our lab, we accommodate the cancer cell line on normal cell line growth media. I hope that BM-MSC cells growth media will be good for leukemia cell line. Otherwise, you can use another growth media that is suitable for your both cell lines.
It will be good to calculate the IC50 values for your cell lines separately before co-culturing.
There are various co-culturing assays. On one hand, you can perform indirect co-culturing using trans-well inserts with different pore size depending on your experiments. On the other hand, you can perform direct co-culture between your cell lines. For direct co-culture, you can use ibidi 2-well inserts too.
If you are looking for microscopy evaluation I suggest staining the cells before starting in the case of direct co-culture. You can use cell membrane staining procedures.
Regarding cells proportions, you can try different ratios 1:2, 1:1, 2:1 or even more combinations.
Thank you, I am happy that you handled the co-culture experiments. Success in your research :) All the best
Dear Manasi Nagare
We have a publication about co-culturing MSCs with myeloma cells. You can have look if you want.
Myeloma cells can corrupt senescent mesenchymal stromal cells and impair their anti-tumor activity October 2015 Oncotarget 6(37) DOI:10.18632/oncotarget.5430
Thank you Mustafa Burak Acar. I will definitely go through the shared article.
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