Generating Oligodendrocyte Precursor Cells

OPCs used in this study were generated from two separate EpiSC lines, EpiSC9 (female) and 129O1 (male), using in vitro differentiation protocols and culture conditions described previously^{3 (link),23}. Cultures were regularly tested and shown to be mycoplasma free. To ensure uniformity throughout all in vitro screening experiments, EpiSC-derived OPCs were sorted to purity by fluorescent activated cell sorting at passage five with conjugated CD140a-APC (eBioscience, 17–1401; 1:80) and NG2-AF488 (Millipore, AB5320A4; 1:100) antibodies. Sorted batches of OPCs were expanded and frozen down in aliquots. OPCs were thawed into growth conditions for one passage prior to use in screening assays.

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Najm F.J., Madhavan M., Zaremba A., Shick E., Karl R.T., Factor D.C., Miller T.E., Nevin Z.S., Kantor C., Sargent A., Quick K.L., Schlatzer D.M., Tang H., Papoian R., Brimacombe K.R., Shen M., Boxer M.B., Jadhav A., Robinson A.P., Podojil J.R., Miller S.D., Miller R.H, & Tesar P.J. (2015). Drug-based modulation of endogenous stem cells promotes functional remyelination in vivo. Nature, 522(7555), 216-220.

Publication 2015

CD140a-APC NG2-AF488

Antibodies Assays Female Frozen Male Mycoplasma

Corresponding Organization :

Other organizations : Case Western Reserve University, University School, PerkinElmer (United States), University of Cincinnati Medical Center, National Center for Advancing Translational Sciences, National Institutes of Health, Northwestern University

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Variable analysis

independent variables

EpiSC line (EpiSC9 female, 129O1 male)
In vitro differentiation protocols and culture conditions

dependent variables

OPC purity

control variables

Mycoplasma testing
Passage number (five) for OPC sorting
Antibodies used for OPC sorting (CD140a-APC, NG2-AF488)
One passage of OPC expansion and freezing before screening assays

controls

No explicit positive or negative controls mentioned

Annotations

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