Stem cell Differentiation media Differentiation of Human iPSC into Human Neuroepithelial 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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Upstream tips
-If STEMdiff™ Neural Induction Medium is received thawed, immediately place at -20°C or aliquot and store at -20°C. Product performance will not be affected.
Protocol tips
-If not used immediately, the cultureware must be sealed to prevent evaporation of the laminin solution (e.g. with Parafilm®) and can be stored at 2 - 8°C for up to 2 weeks after coating. Allow stored coated cultureware to come to room temperature (15 - 25°C) for 30 minutes before plating cells.
-If the cultureware surface is not fully coated by the Matrigel® solution, it should not be used for human ES or iPS cell culture.
- The incubation time may vary when using different cell lines or other non-enzymatic cell dissociation reagents, therefore dissociation should be monitored under the microscope until the optimal time is determined.
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