Dear Colleagues,
I am currently working on a project that involves differentiating human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs) into mesenchymal stem cells (MSCs). I am using laminin-coated plates, mTeSR, and ReleSR media for maintaining the hPSC and iPSC cultures. For MSC derivation and proliferation, I am culturing the cells in alpha DMEM high glucose medium supplemented with fetal bovine serum (FBS), non-essential amino acids (NEAA), ascorbic acid, basic fibroblast growth factor (bFGF), penicillin/streptomycin, dexamethasone, and a ROCK inhibitor. Trypsin is being used to detach the cells from the culture wells.
However, I have encountered some challenges with my existing protocol. Despite trying various culture media and growth factor combinations, my cells fail to proliferate and ultimately die after two days of culture.
Could someone kindly share a reliable and detailed protocol for successfully inducing hESCs or iPSCs to differentiate into MSCs? I would greatly appreciate any guidance or recommendations that could help me overcome the issues I am facing with cell survival and proliferation during the differentiation process.
Obtaining a well-established protocol would be invaluable for my research, and I would be extremely grateful for your assistance and expertise in this matter.
Thank you in advance for your time and consideration.
You might want to compare this protocol to the one you are currently using
Article hiPSC-derived bone marrow milieu identifies a clinically act...
Thank you Dr Sabine Strehl
I checked this article in detail but it is different from my study.
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