I am playing around with media for mesenchymal stem cells isolation from umbilical cord. The standard formula is DMEM (high glucose) + 5% human platelet lysate + 1x Glutamax. Is there any additional stuff I should add to improve/preserve the stemness of the cells? I define stemness as the ability to differentiate highly efficiently to all 3 lineages (Adipo-, Osteo-, and Chondro). Thanks!
Dear Phua,
We used ordinary DMEM, not high glucose. We used lab made plate lysate 10%, but 5% I think will ve OK. However you should pay attention on the timing of harvest. Make it around 80% so the cells do not reach plateau phase yet. If the cells are too confluent, part of them undergo differentiation or senescence and lose their stemness.
Good luck
Dear Kyle,
I suppose you are doing great with 5% platelet lysate. My advice is to use low glucose DMEM. That high glucose media is too high and mimic diabetic environment for healthy cells, their metabolic pathways can be disturbed. But in general, non-immortalised MSCs give best results till 3rd passage even they are able to grow further.
Best
Karolina
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