Stem cell Differentiation media Differentiation of Human iPSCs into Basal Forebrain cholinergic neurons (BFCN)

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

BrainPhys™ Without Phenol Red + mTeSR™1

BrainPhys™ Without Phenol Red

STEMCELL technologies


STEMCELL technologies

Upstream tips
-The seeding density of cells should be optimized for the application and the cell line.
Protocol tips
-Do not prepare the Poly-D-Lysine (PDL) solution with borate buffer. Increased cell clumping is observed when the PDL solution is prepared in borate buffer, and then cells are cultured in medium supplemented with NeuroCult™ SM1.
- Ensure that the coverslips are completely submerged in the PDL solution, as they tend to float. If this happens, use a sterile plastic pipette tip to push the coverslip to the bottom of the well.
Downstream tips
-If not used immediately, aliquot PDL stock solution into polypropylene vials and store at 2 - 8°C for up to 1 month.

A&A Biotechnology

Protocol tips
Shift iPSCs on day 9, gradually to Brainphys media supplemented with B27 (Life Technologies).
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