Generation and Characterization of hiPSCs

Human induced pluripotent stem cells (hiPSCs) were reprogrammed, expanded and characterized for pluripotency and differentiation capacity as described before (29 (link)). In brief, erythroblasts from three donors (age/sex: donor 1: 27/F, donor 2: 50/M, donor 3: 49/F) were reprogrammed by nucleofection of plasmids encoding for OCT4, shRNA-p53, SOX2, KLF4, L-Myc and Lin28. Human iPSCs were cultured using ReproTeSR medium (STEMCELL Technologies, Grenoble, France) and expanded using StemMACS iPSC-Brew XF medium (Miltenyi Biotec, Bergisch Gladbach, Germany). Five hiPSC clones from 3 donors (donor 1: 2 clones, donor 2: 2 clones, donor 3: 1 clone) were used in this study. For some experiments, we also used the IMR90-4 iPSC line (30 ). Human iPSCs were maintained on Matrigel (Corning)-coated plates in mTeSR1 medium (STEMCELL Technologies, Grenoble, France).

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Nishihara H., Gastfriend B.D., Soldati S., Perriot S., Mathias A., Sano Y., Shimizu F., Gosselet F., Kanda T., Palecek S.P., Du Pasquier R., Shusta E.V, & Engelhardt B. (2020). Advancing human induced pluripotent stem cell‐derived blood‐brain barrier models for studying immune cell interactions. The FASEB Journal, 34(12), 16693-16715.

Publication 2020

ReproTeSR medium StemMACS iPSC-Brew XF medium Matrigel mTeSR1 medium

Clone Donor Donors Erythroblasts Hipsc Human Ipscs Klf4 Matrigel Oct4 Plasmids Shrna Sox2 Stemcell

Corresponding Organization : Neurological Surgery

Other organizations : University of Lausanne, Yamaguchi University, Université d'Artois

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Variable analysis

independent variables

Reprogramming method: Nucleofection of plasmids encoding for OCT4, shRNA-p53, SOX2, KLF4, L-Myc and Lin28
Cell type: Erythroblasts from three donors (age/sex: donor 1: 27/F, donor 2: 50/M, donor 3: 49/F)

dependent variables

Pluripotency of hiPSCs
Differentiation capacity of hiPSCs

control variables

Culture conditions: hiPSCs were cultured using ReproTeSR medium, StemMACS iPSC-Brew XF medium, and mTeSR1 medium on Matrigel-coated plates
Cell lines: Five hiPSC clones from 3 donors (donor 1: 2 clones, donor 2: 2 clones, donor 3: 1 clone) and the IMR90-4 iPSC line

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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