Isolation and Culture of iPSC-O Cells

Human cells were isolated from surplus liver tissue of a one-year-old girl with fulminant hepatitis (Hep2064). Primary cells were isolated according to the collagenase perfusion method^{25 (link),29 (link),30 (link)}. iPSC-O cells were generated by introduction of Sendai virus carrying the 4 Yamanaka factors^{31 (link)}. The iPSC-O cells were cultured on MEFs and grown using a medium for human ES cells [KNOCKOUT DMEM (Gibco, 10829), 15% CTS KNOCKOUT SR XenoFree Medium (Gibco, 14150), 1% penicillin streptomycin (Gibco, 11140), 1% non-essential amino acids (Gibco, 11360), 1% Sodium Pyruvate (Gibco, 11360), 1% Gulta MAX (Gibco, 35050), 0.1% β-mercaptoethanol (Gibco, 21985-023), and 10.0 ng/ml bFGF (INVITROGEN, PHG0024)].

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Tsuneishi R., Saku N., Miyata S., Akiyama S., Javaregowda P.K., Ite K., Takashima N., Toyoda M., Kimura T., Kuroda M., Nakazawa A., Kasahara M., Nonaka H., Kamiya A., Kiyono T., Yamauchi J, & Umezawa A. (2021). Ammonia-based enrichment and long-term propagation of zone I hepatocyte-like cells. Scientific Reports, 11, 11381.

Publication 2021

KNOCKOUT DMEM CTS KNOCKOUT SR XenoFree Medium penicillin streptomycin non-essential amino acids Sodium Pyruvate Gulta MAX β-mercaptoethanol bFGF

Cells Collagenase Essential amino acids Fulminant hepatitis Girl Human Human es cells Ipsc Liver Mercaptoethanol Penicillin Perfusion Pyruvate Sendai virus Sodium Streptomycin Tissue

Corresponding Organization :

Other organizations : National Center For Child Health and Development, Kitasato University, Tokyo Medical University, Tokai University, Chiba Cancer Center, Tokyo University of Pharmacy and Life Sciences

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Variable analysis

independent variables

Introduction of Sendai virus carrying the 4 Yamanaka factors to generate iPSC-O cells

dependent variables

Generation of iPSC-O cells from human cells isolated from surplus liver tissue of a one-year-old girl with fulminant hepatitis

control variables

Culture of iPSC-O cells on MEFs
Growth media for human ES cells (KNOCKOUT DMEM, 15% CTS KNOCKOUT SR XenoFree Medium, 1% penicillin streptomycin, 1% non-essential amino acids, 1% Sodium Pyruvate, 1% Gulta MAX, 0.1% β-mercaptoethanol, and 10.0 ng/ml bFGF)

controls

Positive control: Not specified
Negative control: Not specified

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