Stem cell Differentiation media hESCs differentiation into cortical neuroepithelium (NE)

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 3 matching solutions for this experiment

Gibco™Glasgow's MEM (GMEM)

Thermo Fisher Scientific

Protocol tips
cortex differentiation medium (9,000 cells per well) containing Glasgow-MEM, 20% Knockout Serum Replacement, and 20 μM Y-27632. IWR1e (Wnt inhibitor) and SB431542 (TGFβ inhibitor) were added to culture to reach 3 and 5 μM, respectively, from day 0 to day 18
Gibco™DMEM/F-12

Thermo Fisher Scientific

Protocol tips
3N medium consisted of a 1:1 mixture of N2- and B27-containing media. N2 medium: DMEM/F12, N2 (GIBCO), 5 μg/ml Insulin, 1mM L-Glutamine, 100 μm non-essential amino acids, 100 μM 2-mercaptoethanol, 50 U/ml Penicillin and 50 mg/ml Streptomycin; B27 medium: Neurobasal (Invitrogen), B27 with or without vitamin A (GIBCO), 200 mM Glutamine, 50 U/ml Penicillin and 50 mg/ml Streptomycin. 3N medium was supplemented with either 1 μm Dorsomorphin (Tocris) or 500 ng/ml mouse Noggin-CF chimera (R&D Systems), and 10 μm SB431542 (Tocris) to inhibit TGFβ signaling during neural induction
Protocol tips
knock out serum replacement (KSR) media with 10 nM TGF-b inhibitor (SB431542, Tocris) and 500 ng/mL of Noggin (R&D)
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