Generation of Integration-free iPSCs

IPSCs were generated as described^{15 (link)}. Cyto-Tune Sendai reprogramming kit (ThermoFisher) was used according to the manufacturer’s instructions and colonies were expanded on Matrigel-coated (BD Biosciences) plates in mTeSR1 medium (Stemcell Technologies). Lack of genomic integration of Sendai virus into iPSCs was confirmed by PCR and karyotype analysis confirmed the expected chromosomal copy numbers^{16 (link)}. Every 7–8 days, iPSCs were treated with collagenase, and passaged. To verify pluripotency, cells were characterized for Nanog and Tra1–60^{17 (link)}.

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Mishra H.K., Ying N.M., Luis A., Wei H., Nguyen M., Nakhla T., Vandenburgh S., Alda M., Berrettini W.H., Brennand K.J., Calabrese J.R., Coryell W.H., Frye M.A., Gage F.H., Gershon E.S., McInnis M.G., Nievergelt C.M., Nurnberger J.I., Shilling P.D., Oedegaard K.J., Zandi P.P., Kelsoe J.R., Welsh D.K, & McCarthy M.J. (2021). Circadian rhythms in bipolar disorder patient-derived neurons predict lithium response: Preliminary studies. Molecular psychiatry, 26(7), 3383-3394.

Publication 2021

Cyto-Tune Sendai reprogramming kit Matrigel-coated mTeSR1 medium

Cells Chromosomal Collagenase Ipscs Karyotype Matrigel Stemcell Virus genomic

Corresponding Organization : Circadian (United States)

Other organizations : Dalhousie University, University of Pennsylvania, Icahn School of Medicine at Mount Sinai, Case Western Reserve University, University of Iowa, Mayo Clinic, WinnMed, Salk Institute for Biological Studies, University of Chicago, University of Michigan–Ann Arbor, Indiana University – Purdue University Indianapolis, Haukeland University Hospital, University of Bergen, Johns Hopkins University

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Variable analysis

independent variables

Cyto-Tune Sendai reprogramming kit (ThermoFisher)

dependent variables

Lack of genomic integration of Sendai virus into iPSCs
Chromosomal copy numbers
Expression of Nanog and Tra1–60

control variables

Matrigel-coated (BD Biosciences) plates
MTeSR1 medium (Stemcell Technologies)
Passaging of iPSCs every 7–8 days using collagenase

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