BrainPhys™ Without Phenol Red

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-The seeding density of cells should be optimized for the application and the cell line.	-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.	-If not used immediately, aliquot PDL stock solution into polypropylene vials and store at 2 - 8°C for up to 1 month.

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.

Publication protocol

7889(s)B, 050643 (Control), 948 (AD1), 949(fControl), and 950 (AD2) iPSC lines were obtained via the NYSCF Repository following the guidelines from [60]. The derivation and characterization of Nkx2.1-GFP ESC line was previously published [30]. ES and iPS cell lines were expanded and maintained in serum-free mTeSR1 media (Stem Cell Technologies). Cells were lifted using StremPro Accutase (ThermoFisher) and media was supplemented with 10 μM ROCK inhibitor (Y27632, Stemgent) during cell passaging.
For all studies in this paper, cell lines underwent at least 3 independent differentiations from the iPSC stage to the mature neuron stage. Data were routinely compared across these independently derived genotype-identical neurons (or in some cases neuronal precursors), and if comparable results were obtained across independently genotype-identical derived cells, they were considered to be qualified representatives of their genotype and so were passed along for genotype-specific experimentation.

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Discussion

Papers

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Manufacturer protocol

Download the product protocol from STEMCELL technologies for BrainPhys™ Without Phenol Red below.

Download manufacturer protocol

Videos

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