Integration-free iPSC-derived NSPC protocols

Three lines of integration free human PBMC-derived iPSCs (1210B2, 1231A3, and 1201C1), which were established from ePBMCs® from the Cellular Technology Limited (OH, USA) at Center for iPS Cell Research and Application (CiRA: Kyoto, Japan) by an integration-free method [51 (link)], were used. The 1210B2 and 1231A3 iPSCs were cultured with a feeder-free protocol [33 (link)], and the 1201C1 iPSCs were cultured with an on-feeder protocol that uses SNL feeder cells. They were induced into NSPCs as previously described [52 (link), 53 (link)] with two slight modifications. Briefly, in the first protocol, the NSPCs were induced directly from embryoid bodies (EBs) by a protocol that consists only of a floating culture. In the second protocol, the EBs were adhered to laminin-coated culture dishes on day 7, and they subsequently formed neural rosettes (NRs), which were picked on day 14. We refer to NSPCs induced directly from EBs as EB-NSPCs, and those induced from the NR phase as NR-NSPCs. The NSPCs were expanded using the neurosphere culture technique [23 (link), 54 (link)].

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Sugai K., Fukuzawa R., Shofuda T., Fukusumi H., Kawabata S., Nishiyama Y., Higuchi Y., Kawai K., Isoda M., Kanematsu D., Hashimoto-Tamaoki T., Kohyama J., Iwanami A., Suemizu H., Ikeda E., Matsumoto M., Kanemura Y., Nakamura M, & Okano H. (2016). Pathological classification of human iPSC-derived neural stem/progenitor cells towards safety assessment of transplantation therapy for CNS diseases. Molecular Brain, 9, 85.

Publication 2016

ePBMCs

Cellular Culture technique Dishes Embryoid bodies Feeder cells Human Ipscs Laminin Neural

Corresponding Organization :

Other organizations : Keio University, Tokyo Metropolitan Children's Medical Center, Osaka National Hospital, Keio University Hospital, Central Institute for Experimental Animals, Hyogo Medical University, Yamaguchi University

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Variable analysis

independent variables

Induction method for NSPCs (direct from EBs vs. from NRs)

dependent variables

NSPC characteristics and properties

control variables

IPSC lines (1210B2, 1231A3, and 1201C1)
Feeder-free vs. on-feeder culture protocols for iPSCs

controls

Positive control: None explicitly mentioned
Negative control: None explicitly mentioned

Annotations

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