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Stem Cell Reports. 2017 Apr 11;8(4):1101-1111. doi: 10.1016/j.stemcr.2017.03.011.

High-Throughput and Cost-Effective Characterization of Induced Pluripotent Stem Cells.

Author information

1
Department of Pediatrics, University of California, San Diego, La Jolla, CA 92093, USA.
2
Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA.
3
Institute for Genomic Medicine, University of California, San Diego, La Jolla, CA 92093, USA.
4
Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA; Institute for Genomic Medicine, University of California, San Diego, La Jolla, CA 92093, USA.
5
Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA; Department of Neurosciences, University of California, San Diego, La Jolla, CA 92093, USA. Electronic address: lgoldstein@ucsd.edu.
6
Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA. Electronic address: apanopou@nd.edu.
7
Department of Pediatrics, University of California, San Diego, La Jolla, CA 92093, USA; Institute for Genomic Medicine, University of California, San Diego, La Jolla, CA 92093, USA. Electronic address: kafrazer@ucsd.edu.

Abstract

Reprogramming somatic cells to induced pluripotent stem cells (iPSCs) offers the possibility of studying the molecular mechanisms underlying human diseases in cell types difficult to extract from living patients, such as neurons and cardiomyocytes. To date, studies have been published that use small panels of iPSC-derived cell lines to study monogenic diseases. However, to study complex diseases, where the genetic variation underlying the disorder is unknown, a sizable number of patient-specific iPSC lines and controls need to be generated. Currently the methods for deriving and characterizing iPSCs are time consuming, expensive, and, in some cases, descriptive but not quantitative. Here we set out to develop a set of simple methods that reduce cost and increase throughput in the characterization of iPSC lines. Specifically, we outline methods for high-throughput quantification of surface markers, gene expression analysis of in vitro differentiation potential, and evaluation of karyotype with markedly reduced cost.

KEYWORDS:

SNP arrays; differentiation potential; digital karyotyping; flow cytometry; fluorescent cell barcoding; high-throughput methods; induced pluripotent stem cells; pluripotency characterization; qPCR

PMID:
28410643
PMCID:
PMC5390243
DOI:
10.1016/j.stemcr.2017.03.011
[Indexed for MEDLINE]
Free PMC Article

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