Stem cell Differentiation media hiPSCs or hESCs 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 (GSK3š›ƒ-i, TGFš›ƒ-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-š›ƒ3) 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 5 matching solutions for this experiment

Gibcoā„¢KnockOutā„¢ DMEM

Thermo Fisher Scientific

Upstream tips
growth media (80% KO-DMEM, 20% Knock-out Serum Replacer (KSR), 1% non-essential amino acids, 2 mM Glutamax, 0.1 mM Ɵ-Mercaptoethanol, and 1% penicillin and streptomycin all Life Technologies), supplemented with either 10 ng/ml (Shef hES and NAS2 hiPS) or 20 ng/ml (MSU0001) FGF2 (Peprotech).
Protocol tips
Colonies were chopped into uniform 150 Ī¼m pieces using a McIlwain tissue chopper (Mickle Engineering, Gomshall, U.K.), and resuspended in growth media (described above), without FGF2 but supplemented with 10 Ī¼M of the ROCK inhibitor Y27632 (Tocris)
Downstream tips
Colony pieces formed spheres and differentiated into EBs over 4 subsequent days, after which they were transferred into modified chemically defined media (CDM) (Joannides et al., 2007), (BSA was substituted with 400 Ī¼g/ml Albumax-II, Life Technologies). CDM was supplemented with 10 Ī¼M Y27632 (Tocris) and 20 Ī¼M of the Nodal/TGF-Ī² signaling inhibitor SB431542 (Tocris)
Upstream tips
Growth Medium: DMEM/F12 (Merck Millipore, Germany) supplemented with 20% KnockOutā„¢ Serum Replacement (KnockOutā„¢ SR, Thermo Fisher Scientific Inc., MA, USA), 1% penicillin-streptomycin (Sigma-Aldrich; Merck KGaA), 1 mM non-essential amino acid (NEAA; Sigma-Aldrich; Merck KGaA), 0.2 mM 2-mercaptoethanol (Sigma-Aldrich; Merck KGaA), and 10 ng/ml basic fibroblast growth factor (bFGF) (Sigma-Aldrich; Merck KGaA)
Protocol tips
Growth medium with the addition of 10 ĀµM ROCK-inhibitor Y-27632 (Sigma-Aldrich; Merck KGaA). Next, cells were left for spontaneous aggregation for 15 days. The medium (hESC medium without bFGF) was changed every second day.
Gibcoā„¢IMDM

Thermo Fisher Scientific

Upstream tips
human pluripotent stem cells were detached from the feeder layer by collagenase IV, dispersed into small clumps
Protocol tips
Iscoveā€™s modified Dulbeccoā€™s differentiation medium supplemented with 20% fetal calf serum, 1Ā mMĀ l-glutamine, 0.1Ā mM beta-mercaptoethanol and 1% nonessential amino acid stock in ultra-low attachment plates (Corning) for 7Ā days.
Protocol tips
The cells were centrifuged at 300g for 3 min and then resuspended in Apelā„¢ media (Stem Cell Technologies) containing 10 Ī¼M Rock inhibitor at 30,000 cells per ml. 100 Ī¼l was then added to each well of a U-bottom 96 well plate and centrifuged at 300g for 3 min.
Gibcoā„¢ StemProā„¢ hESC SFM

Thermo Fisher Scientific

Protocol tips
Cell aggregates were suspended in EB formation medium (50% DMEM/F12 (Life Technologies, 11330-032) 50% mTeSRā„¢1 or StemProĀ® hESC SFM containing 10 ĀµM Rock Inhibitor Y27632 (Millipore, SCM075)) and allowed to settle by gravity in a conical tube.
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