Culturing Human Embryonic Stem Cells and Derived Cell Lines

Human H9 ESCs as well as derived YAP^−/− and TAZ^−/− hESCs were maintained on feeder layers of mitomycin C–inactivated MEFs in hESC medium [68 (link)] (DMEM/F12 [Thermo Fisher Scientific, Waltham, MA], 20% Knockout Serum Replacement [Thermo Fisher Scientific], 0.1 mM nonessential amino acids [NEAAs; Thermo Fisher Scientific], 2 mM GlutaMAX [Thermo Fisher Scientific], 1% penicillin/streptomycin [Thermo Fisher Scientific], 55 μM β-mercaptoethanol [Thermo Fisher Scientific], and 10 ng/ml bFGF [Joint Protein Central, Incheon, Korea]) or on Matrigel (BD Biosciences, San Jose, CA, USA) in mTeSR medium (STEMCELL Technologies, Vancouver, Canada). hESCs derived hMSCs and BM-hMSCs (purchased from Lonza, Basel, Switzerland) were cultured in hMSC medium (αMEM + GlutaMAX [Thermo Fisher Scientific], 10% fetal bovine serum [Gibco, Cat: 10099–141, Lot: 1616964], 1% penicillin/streptomycin [Thermo Fisher Scientific], and 1 ng/ml bFGF [Joint Protein Central]). No mycoplasma contamination was observed during cell culture.

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Fu L., Hu Y., Song M., Liu Z., Zhang W., Yu F.X., Wu J., Wang S., Izpisua Belmonte J.C., Chan P., Qu J., Tang F, & Liu G.H. (2019). Up-regulation of FOXD1 by YAP alleviates senescence and osteoarthritis. PLoS Biology, 17(4), e3000201.

Publication 2019

DMEM/F12 Knockout Serum Replacement NEAAs GlutaMAX penicillin/streptomycin β-mercaptoethanol Matrigel mTeSR medium BM-hMSCs αMEM + GlutaMAX fetal bovine serum

Amino acids Cell culture Cultured medium Escs Feeder layers Fetal bovine serum Hescs Human Joint Matrigel Mercaptoethanol Mitomycin c Mycoplasma Penicillin Protein Serum Stemcell Streptomycin

Corresponding Organization : Chinese Institute for Brain Research

Other organizations : Children's Hospital of Fudan University, The University of Texas Southwestern Medical Center, Salk Institute for Biological Studies

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Variable analysis

independent variables

Genetic manipulation (YAP−/− and TAZ−/− hESCs)

dependent variables

Not explicitly mentioned

control variables

Human H9 ESCs (wild-type)
Culture conditions (feeder layers of mitomycin C–inactivated MEFs in hESC medium or on Matrigel in mTeSR medium)
HMSC culture conditions (αMEM + GlutaMAX, 10% fetal bovine serum, 1% penicillin/streptomycin, and 1 ng/ml bFGF)

controls

Positive control: Human H9 ESCs (wild-type)
Negative control: Not mentioned

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