Stem cell Differentiation media mPSCs/mESCs differentiation to Embryoid body (EB)

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™KnockOut™ DMEM

Thermo Fisher Scientific

Upstream tips
20 000 cells per 200 μl in a V-shaped well of a 96-well plate (Sumitomo Bakelite, Tokyo, Japan) with an ultra-low-cell adhesion surface
Protocol tips
KnockOut DMEM (Life Technologies, Carlsbad, CA, USA) containing 15% KnockOut Serum Replacement (HyClone, Logan, UT, USA), 1% GlutaMAX (Life Technologies), 1% MEM Non-Essential Amino Acids (Life Technologies), 0.09% 2-mercaptoethanol (Life Technologies), and 0.1% Leukemia Inhibitory Factor (Wako, Osaka, Japan)
Gibco™Glasgow's MEM (GMEM)

Thermo Fisher Scientific

Protocol tips
differentiation medium (G-MEM (Gibco #11710-035), supplemented with 10% KnockOut™ Serum Replacement (Gibco #10828-028, Lot 1667387), 1% non-essential amino acids (GE Healthcare), 0.1 mM β-mercaptoethanol (Sigma Aldrich), 200 U/ml penicillin (Sigma Aldrich), 200 μg/ml streptomycin (Sigma Aldrich)) per 1/96 well (Nunclon Sphera® Low Adhesion Plate 96-U, Thermo Scientific #174925).
Downstream tips
On day six, aggregates were transferred to 90 mm plates with low adhesion surface (Nunclon Sphera® 90 mm, Thermo Scientific #174945) and subsequently cultured in N2 medium (49% DMEM low glucose (1 g/l, GE Healthcare #E15-005), 49% HAM's F12 (GE Healthcare # E15-016), 1 × N2 supplement (100 ×, Gibco #17502-048 Lot 1737260) and 200 U/ml penicillin (Sigma Aldrich), 200 μg/ml streptomycin (Sigma Aldrich)).
Gibco™DMEM/F-12

Thermo Fisher Scientific

Upstream tips
cells were transferred to low adhesion tissue culture plates coated with 2% poly-HEMA in ethanol.
Protocol tips
EBM [DMEM-F12, 15% Knock Out Serum Replacement (KOSR), 15% HI FBS, 1 mM L-glutamine, 0.1 mM 2-mercaptoethanol, and 1% Non-essential amino acids (NEAA, Invitrogen, Grand Island, NY), 1% antibiotic-antimycotic solution (Invitrogen), and 40 ng/ml human Beta fibroblast growth factor (β-FGF)] in suspension for a minimum of 14 days, during which they formed EBs.
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