Stem cell Differentiation media hPSC or hiPSCS differentiation into myogenic cells

Stem cells have the unique ability to self-renew or differentiate themselves into various cell types in response to appropriate signals. These cells are especially important for tissue repair, regeneration, replacement, or in the case of hematopoietic stem cells (HSCs) to differentiate into various myeloid populations. Appropriate signals refer to the growth factor supplements or cytokines that mediate differentiation of various stem cells into the required differentiated form. For instance, HSCs can be differentiated into dendritic cells (with IL-4 and GM-CSF), macrophages (with m-CSF) and MDSCs (with IL-6 and GM-CSF). Human pluripotent stem cells (hPSCs) and induced pluripotent stem cells (iPSCs) can be first cultured in neural differentiation media (GSK3𝛃-i, TGF𝛃-i, AMPK-i, hLIF) to form neural rosettes, which can be differentiated into neural or glial progenitors (finally differentiated into oligodendrocytes). Neural progenitors can be finally differentiated into glutaminergic (dibytyryl cAMP, ascorbic acid) and dopaminergic (SHH, FGF-8, BDNF, GDNF, TGF-𝛃3) neurons. Thus, it is important to first identify the self-renewing cell line: its source and its final differentiation state, followed by the supplements and cytokines required for the differentiation, and final use. Timelines are another thing that is considered. For instance, it takes 7-10 days to form neural rosettes from iPSCs and 3 days to differentiate neural progenitors to neurons. Finally, the stability for stem cell culture media varies. It is advised to make fresh media every time when differentiating HSCs to myeloid populations, whereas neural differentiation media may remain stable for two weeks when stored in dark between 2-8C.

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Found 2 matching solutions for this experiment

Gibco™Essential 8™ Medium

Thermo Fisher Scientific

Upstream tips
Prior to differentiation, it is essential to ensure 80–90% of cells are undifferentiated and cultures are 70–80% confluent. Prior to dissociation cells are treated with 10 μM ROCK inhibitor (Y-27632) to reduce dissociation-induced apoptosis
Protocol tips
E8 media supplemented with 10 μM Y-27632
Downstream tips
Cells are then treated with 10 μM CHIR99021 supplemented E6 media—E8 media lacking the pluripotency factors FGF2 and TGF-β1—to simulate Wnt signaling during gastrulation
TeSR™-E8™

STEMCELL technologies

Upstream tips
cells infected with Tet-on lentivirus
Protocol tips
E8 medium supplemented with Y27632 (5 µM, Tocris) at 3.3 × 104 cells/cm2. The following day, E8 media was replaced with E6 media and cells were cultured for 2 days supplemented with CHIR99021 (10 µM, Selleck Chemical), after which CHIR99021 was removed and E6 media supplemented with 1 µg/mL Dox (Sigma) for 18 days until GFP+ induced myogenic progenitor cells (iMPCs) were sorted by FACS as described below. During differentiation of iMPCs, 10 ng/mL bFGF (R&D) was added starting at day 5 to enhance proliferation of GFP+ cells.
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