Culturing Primed Human Embryonic Stem Cells

UCLA2 and UCLA1 hESC are cultured as previously described^{16 (link)}, briefly the hESCs are cultured in hESC media, which was composed of 20% knockout serum replacement (KSR) (Life Technologies, A3181502), 1x MEM Non-Essential Amino Acids (NEAA) (Fisher Scientific, 25-025-CI), 1x Penicillin/Streptomycin/Glutamine (Thermo Fisher, 10378016), 55 µM 2- Mercaptoethanol (Life Technologies, 21985-023), 10 ng/mL recombinant human FGF basic (Proteintech, HZ1285), and 50 ng/mL primocin (InvivoGen, ant-pm-2) in DMEM/F12 media (GIBCO, 11330-032). The primed hESCs were split by 1 mg/ml Collagenase type IV (GIBCO, 17104-019) and maintained routinely on mitomycin C (MMC)-inactivated mouse embryonic fibroblasts (MEFs). The hESCs were split every 7 days using Collagenase type IV (GIBCO, 17104-019). HEK293 cells were acquired from ATCC (Cat# CRL-3216). No lines used in this study belong to the International Cell Line Authentication Committee register of misidentified cell lines.

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Xiang X., Tao Y., DiRusso J., Hsu F.M., Zhang J., Xue Z., Pontis J., Trono D., Liu W, & Clark A.T. (2022). Human reproduction is regulated by retrotransposons derived from ancient Hominidae-specific viral infections. Nature Communications, 13, 463.

Publication 2022

Penicillin/Streptomycin/Glutamine 2- Mercaptoethanol recombinant human FGF basic primocin ant-pm-2 Collagenase type IV HEK293 cells

Cell line authentication Collagenase Cultured media Embryonic Essential amino acids Fibroblasts Glutamine Hek293 cells Hescs Human Mercaptoethanol Mitomycin c Mouse Penicillin Serum Streptomycin Type iv collagenase

Corresponding Organization : Second Affiliated Hospital of Zhejiang University

Other organizations : Zhejiang University-University of Edinburgh Institute, University of California, Los Angeles, École Polytechnique Fédérale de Lausanne

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Variable analysis

independent variables

Culture protocol (UCLA2 and UCLA1 hESC)

dependent variables

Not explicitly mentioned

control variables

HESC media composition (20% knockout serum replacement, 1x MEM Non-Essential Amino Acids, 1x Penicillin/Streptomycin/Glutamine, 55 µM 2-Mercaptoethanol, 10 ng/mL recombinant human FGF basic, 50 ng/mL primocin in DMEM/F12 media)
Culture conditions (maintained on mitomycin C (MMC)-inactivated mouse embryonic fibroblasts (MEFs))
Cell line identity (No lines used in this study belong to the International Cell Line Authentication Committee register of misidentified cell lines)

positive controls

Not specified

negative controls

Not specified

Annotations

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