Generation of integration-free human induced pluripotent stem cells from postnatal blood mononuclear cells by plasmid vector expression

Sarah N. Dowey, Xiaosong Huang, Bin Kuan Chou, Zhaohui Ye, Linzhao Cheng

Research output: Contribution to journalArticle

Abstract

Several human postnatal somatic cell types have been successfully reprogrammed to induced pluripotent stem cells (iPSCs). Blood mononuclear cells (MNCs) offer several advantages compared with other cell types. They are easily isolated from umbilical cord blood (CB) or adult peripheral blood (PB), and can be used fresh or after freezing. A short culture allows for more efficient reprogramming, with iPSC colonies forming from blood MNCs in 14 d, compared with 28 d for age-matched fibroblastic cells. The advantages of briefly cultured blood MNCs may be due to favorable epigenetic profiles and gene expression patterns. Blood cells from adults, especially nonlymphoid cells that are replenished frequently from intermittently activated blood stem cells, are short-lived in vivo and may contain less somatic mutations than skin fibroblasts, which are more exposed to environmental mutagens over time. We describe here a detailed, validated protocol for effective generation of integration-free human iPSCs from blood MNCs by plasmid vectors.

Original languageEnglish (US)
Pages (from-to)2013-2021
Number of pages9
JournalNature Protocols
Volume7
Issue number11
DOIs
StatePublished - Nov 2012

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Induced Pluripotent Stem Cells
Stem cells
Blood Cells
Plasmids
Blood
Mutagens
Fetal Blood
Transcriptome
Epigenomics
Freezing
Cultured Cells
Stem Cells
Fibroblasts
Cell culture
Gene expression
Skin
Mutation
Cells

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Generation of integration-free human induced pluripotent stem cells from postnatal blood mononuclear cells by plasmid vector expression. / Dowey, Sarah N.; Huang, Xiaosong; Chou, Bin Kuan; Ye, Zhaohui; Cheng, Linzhao.

In: Nature Protocols, Vol. 7, No. 11, 11.2012, p. 2013-2021.

Research output: Contribution to journalArticle

Dowey, Sarah N. ; Huang, Xiaosong ; Chou, Bin Kuan ; Ye, Zhaohui ; Cheng, Linzhao. / Generation of integration-free human induced pluripotent stem cells from postnatal blood mononuclear cells by plasmid vector expression. In: Nature Protocols. 2012 ; Vol. 7, No. 11. pp. 2013-2021.
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