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Methods Mol Biol. 2016;1353:285-307. doi: 10.1007/7651_2015_205.

Generation of Human Induced Pluripotent Stem Cells Using RNA-Based Sendai Virus System and Pluripotency Validation of the Resulting Cell Population.

Author information

1
Institute of Genetic Medicine, Newcastle University, Newcastle-upon-Tyne, UK. v.chichagova@newcastle.ac.uk.
2
Institute of Genetic Medicine, Newcastle University, Newcastle-upon-Tyne, UK.
3
Sunderland Eye Infirmary, Sunderland, UK.

Abstract

Human induced pluripotent stem cells (hiPSCs) provide a platform for studying human disease in vitro, increase our understanding of human embryonic development, and provide clinically relevant cell types for transplantation, drug testing, and toxicology studies. Since their discovery, numerous advances have been made in order to eliminate issues such as vector integration into the host genome, low reprogramming efficiency, incomplete reprogramming and acquisition of genomic instabilities. One of the ways to achieve integration-free reprogramming is by using RNA-based Sendai virus. Here we describe a method to generate hiPSCs with Sendai virus in both feeder-free and feeder-dependent culture systems. Additionally, we illustrate methods by which to validate pluripotency of the resulting stem cell population.

KEYWORDS:

Pluripotency; Sendai; hiPSC

PMID:
25697416
DOI:
10.1007/7651_2015_205
[Indexed for MEDLINE]

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