Detailed Characterization of Human Induced Pluripotent Stem Cells Manufactured for Therapeutic Applications

We have recently described manufacturing of human induced pluripotent stem cells (iPSC) master cell banks (MCB) generated by a clinically compliant process using cord blood as a starting material (Baghbaderani et al. in Stem Cell Reports, 5(4), 647-659, 2015). In this manuscript, we describe the det...

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Bibliographic Details
Published in:Stem cell reviews and reports 2016-08, Vol.12 (4), p.394-420
Main Authors: Baghbaderani, Behnam Ahmadian, Syama, Adhikarla, Sivapatham, Renuka, Pei, Ying, Mukherjee, Odity, Fellner, Thomas, Zeng, Xianmin, Rao, Mahendra S
Format: Article
Language:English
Subjects:
Antigens, CD34 - blood
Cell Differentiation
Cell Line
Cell Proliferation
Cells, Cultured
Comparative Genomic Hybridization - methods
Embryoid Bodies - cytology
Embryoid Bodies - metabolism
Fetal Blood - cytology
Fetal Blood - metabolism
Flow Cytometry
Gene Expression
Genome, Human - genetics
Humans
Immunohistochemistry
Induced Pluripotent Stem Cells - cytology
Induced Pluripotent Stem Cells - metabolism
Karyotype
Pluripotent Stem Cells - cytology
Pluripotent Stem Cells - metabolism
Polymorphism, Single Nucleotide
Sequence Analysis, DNA - methods
Stem Cell Transplantation - methods
Tissue Banks
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Summary:We have recently described manufacturing of human induced pluripotent stem cells (iPSC) master cell banks (MCB) generated by a clinically compliant process using cord blood as a starting material (Baghbaderani et al. in Stem Cell Reports, 5(4), 647-659, 2015). In this manuscript, we describe the detailed characterization of the two iPSC clones generated using this process, including whole genome sequencing (WGS), microarray, and comparative genomic hybridization (aCGH) single nucleotide polymorphism (SNP) analysis. We compare their profiles with a proposed calibration material and with a reporter subclone and lines made by a similar process from different donors. We believe that iPSCs are likely to be used to make multiple clinical products. We further believe that the lines used as input material will be used at different sites and, given their immortal status, will be used for many years or even decades. Therefore, it will be important to develop assays to monitor the state of the cells and their drift in culture. We suggest that a detailed characterization of the initial status of the cells, a comparison with some calibration material and the development of reporter sublcones will help determine which set of tests will be most useful in monitoring the cells and establishing criteria for discarding a line.
ISSN:2629-3269
2629-3277
DOI:10.1007/s12015-016-9662-8