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 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.
Lonza Walkersville, Inc., Walkersville, MD, 21793, USA.
Behnam Ahmadian
BA
Baghbaderani
Centre for Brain development and Repair, Institute of Stem Cells and Regenerative Medicine (InSTEM), Bangalore, India.
Adhikarla
A
Syama
Buck Institute for Researching on Aging, Novato, CA, USA.
Renuka
R
Sivapatham
XCell Science, Novato, CA, USA.
Ying
Y
Pei
Centre for Brain development and Repair, Institute of Stem Cells and Regenerative Medicine (InSTEM), Bangalore, India.
Odity
O
Mukherjee
Lonza Walkersville, Inc., Walkersville, MD, 21793, USA.
Thomas
T
Fellner
Buck Institute for Researching on Aging, Novato, CA, USA.|XCell Science, Novato, CA, USA.
Xianmin
X
Zeng
NxCell Inc, Novato, CA, USA. mrao@qthera.com.|Q therapeutics, Salt Lake City, UT, USA. mrao@qthera.com.
Mahendra S
MS
Rao
D018952:Antigens, CD34 / Q000097:blood; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D055028:Comparative Genomic Hybridization / Q000379:methods; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D015870:Gene Expression; D015894:Genome, Human / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D059785:Karyotype; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA / Q000379:methods; D033581:Stem Cell Transplantation / Q000379:methods*; D014015:Tissue Banks
D018952:Antigens, CD34
Consent; Embryonic stem cells; Induced pluripotent stem cells; Manufacturing; Markers; cGMP