Differentiation of hESCs to NPCs and MES

H9 hESCs (WiCell) were maintained in mTeSR1 medium (Stem Cell Technologies) on Matrigel (BD Biosciences)-coated tissue culture plates. H9 cells were passaged every 5 days by ReLeSR (Stem Cell Technologies). For NPC differentiation, hESCs were passaged as single cells using Accutase (Life Technologies) on Matrigel with ROCK inhibitor (Y-27632, Millipore) and induced to differentiate with SB431542 (10 µM, Tocris Bioscience) and NOGGIN (200 ng/ml, PeproTech)^{52 (link)}. To derive MES, hESCs were plated on Matrigel-coated plates as single cells and induced to differentiate with mTeSR1 containing 5 ng/ml hBMP4 (R&D)^{59 (link)}.

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Golkaram M., Jang J., Hellander S., Kosik K.S, & Petzold L.R. (2017). The Role of Chromatin Density in Cell Population Heterogeneity during Stem Cell Differentiation. Scientific Reports, 7, 13307.

Publication 2017

H9 hESCs mTeSR1 medium Matrigel ReLeSR Accutase SB431542 NOGGIN hBMP4

Accutase Cells Hescs Matrigel Noggin Sb431542 Stem cell Tissue Y 27632

Corresponding Organization :

Other organizations : University of California, Santa Barbara

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Variable analysis

independent variables

SB431542 (10 µM, Tocris Bioscience)
NOGGIN (200 ng/ml, PeproTech)
HBMP4 (5 ng/ml, R&D)

dependent variables

NPC differentiation
MES differentiation

control variables

H9 hESCs (WiCell) maintained in mTeSR1 medium (Stem Cell Technologies) on Matrigel (BD Biosciences)-coated tissue culture plates
H9 cells passaged every 5 days by ReLeSR (Stem Cell Technologies)
HESCs passaged as single cells using Accutase (Life Technologies) on Matrigel with ROCK inhibitor (Y-27632, Millipore)

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