Im going to co culture CD4 naive cells with mesenchymal stem cells and will add T-helper 17 differentiation media. After 10 days, I will use the cells for further analysis. Do you think the mesenchymal stem cells could survive for 10 days without FBS? If not, could anyone provide me with a trusted protocol.
Does the co culture media contains growth factors? If yes, MSC might survive.
MSCs can survive in PBS for 72 hours, and in Phisiologic saline, and DMREM-HG for 96 hours (viability >70%) when checked by Trypan blue exclusion method.
However, their capacity to attach and proliferate are reduced faster, though they are viable
You need to check the composition of differentiation media. Or before the experiment, you may check the effect of this media on MSCs by culturing just a few wells of the culture plate. If nothing goes wrong you may proceed as required.
You may also first differentiate the CD4 naive cells and then add to MSCs culture.
I have cultured MSCs in serum free media differentiation media for 10 days and they grow well. But I dont know what will be the effect of CD4 cells on it. It survives well in MEM and DMEM serum free media (proliferation gets slower).
Hope it helps
I have cultured MSCs from umbilical cord matrix in serum-free aMEM for 3 days. They did not show any signs of damage (though showed no signs of proliferation). But, after seeding, they had nearly 24 hours in complete media (to "adapt"), and only after that the media was changed. And, I guess, the initial planting density was sufficient (like, 50% confluence after 24 hours). As far as I know, some of MSCs can be sensitive to this.
What is the medium composition you will use? If you just use a basal medium without serum cells will not survive. I cultured MSCs in a chemically defined serum free medium of different companies. MSCs did not grow as well as with serum but survived and were viable. So if you want to cultivate serum-free check for some serum-free but chemically defined media that were developed for cultivation of MSCs.
MSC survives only limited period even if they look O.K. However, the normal-looking cell morphology does not guarantee its normal functionality. Real meaning of it is cell arrest for a brief time until it gets re-supply of all the goodies of serum to proliferate, survive, and interact with other cells and ECM. For the reference, serum-exposed cells could undergo cellular activities about up to 18h in vitro without serum. It is a simple fasting condition for man. He has to eat to survive and maintain. That man consists of body cells!!!
See also the link.
https://www.researchgate.net/post/What_is_the_reason_for_better_translation_of_MSCs_in_serum_conditions_in_culture_rather_than_serum-free?tpr_view=yccvao0mIQ8Q1Sv5RdvMj6AFD7J27yAWWkpA_2
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