Characterization of Human Stem Cell Lines

The human pluripotent stem cell-lines used in this study were obtained with full Ethical/Institutional Review Board approval by the University of Edinburgh and validated using standard methods including chromosomal analysis, pluripotency and absence of plasmid integration. The iPSC lines CS02iCTR-NTn1 (hPSC1, male) and CS25iCTRL-18n2 (hPSC2, male) were obtained from Cedars-Sinai. For the Nfasc155^-/- lines, fibroblasts were obtained by RS with ethical approval granted by the Institutional Review Board of Warsaw Medical University (Smigiel et al., 2018 (link)). The CS00iNK-n1 (Nfasc155^-/-_{clone 1}, female) and CS00iNK-n2 (Nfasc155^-/-_{clone 2}, female) iPSC lines were generated by Cedars-Sinai. The human embryonic stem cell-line SHEF4 (hPSC3, male) was obtained from the UK Stem Cell Bank. iPSCs were maintained on Matrigel (Scientific Laboratory Supplies)-coated 6-well plates in Essential 8 medium (Thermo Fisher Scientific) at 37 °C and 5% CO2. iPSC colonies were passaged by incubating in Dispase (0.5 mg/ml, Thermo Fisher Scientific)/Collagenase (1 mg/ml, Thermo Fisher Scientific) for 15-25 minutes at 37 °C, washed in DPBS and resuspended in Essential 8 medium before being redistributed in new 6-well plates. Cultures were regularly tested and maintained mycoplasma free. These cell-lines have not been authenticated.

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James O.G., Selvaraj B.T., Magnani D., Burr K., Connick P., Barton S.K., Vasistha N.A., Hampton D.W., Story D., Smigiel R., Ploski R., Brophy P.J., ffrench-Constant C., Lyons D.A, & Chandran S. (2021). iPSC-derived myelinoids to study myelin biology of humans. Developmental Cell, 56(9), 1346-1358.e6.

Publication 2021

Matrigel Essential 8 medium Dispase Collagenase

Cell lines Chromosomal Clone Collagenase Dispase Female Fibroblasts Human Human embryonic stem cell Institutional review board Ipsc Male Matrigel Mycoplasma Ntn1 Plasmid Stem Stem cell

Corresponding Organization : Institute for Stem Cell Biology and Regenerative Medicine

Other organizations : Florey Institute of Neuroscience and Mental Health, University of Melbourne, Wroclaw Medical University, Medical University of Warsaw

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Variable analysis

independent variables

Cell lines used: CS02iCTR-NTn1 (hPSC1, male), CS25iCTRL-18n2 (hPSC2, male), CS00iNK-n1 (Nfasc155-/- clone 1, female), CS00iNK-n2 (Nfasc155-/- clone 2, female), and SHEF4 (hPSC3, male)

dependent variables

Outcome measures not explicitly mentioned

control variables

Cell culture conditions: iPSCs maintained on Matrigel-coated 6-well plates in Essential 8 medium at 37 °C and 5% CO2
Cell line validation: Chromosomal analysis, pluripotency, and absence of plasmid integration
Mycoplasma testing: Cultures regularly tested and maintained mycoplasma free

positive controls

None specified

negative controls

None specified

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