Long-term maintenance of undifferentiated human embryonic and induced pluripotent stem cells in suspension.

Abstract:

:Traditionally, undifferentiated pluripotent human embryonic and induced pluripotent stem cells (hESCs and hiPSCs) have been expanded as monolayer colonies in adhesion culture, both in the presence or absence of feeder cells. However, the use of pluripotent stem cells poses the need to scale-up current culture methods. Herein, we present the cultivation of 2 hESC lines (Royan H5 and Royan H6) and 2 hiPSC lines (hiPSC1 and hiPSC4) as carrier-free suspension aggregates for an extended period of time. The cells proliferated over multiple passages kept a stable karyotype, which successfully maintained an undifferentiated state and pluripotency, as determined by marker expressions in addition to in vitro spontaneous and directed differentiation. Additionally, these cells can be easily frozen and thawed without losing their proliferation, karyotype stability, and developmental potential. Transcriptome analysis of the 3 lines revealed that the adherent culture condition was nearly identical to the suspension culture in Royan H5 and hiPSC1, but not in Royan H6. It remains unclear whether this observation at the transcript level is biologically significant. In comparison with recent reports, our study presents a low-cost procedure for long-term suspension expansion of hESCs and hiPSCs with the capability of freeze/thawing, karyotype stability, and pluripotency. Our results will pave the way for scaled up expansion and controlled differentiation of hESCs and hiPSCs needed for cell therapy, research, and industrial applications in a bioreactor culture system.

journal_name

Stem Cells Dev

authors

Larijani MR,Seifinejad A,Pournasr B,Hajihoseini V,Hassani SN,Totonchi M,Yousefi M,Shamsi F,Salekdeh GH,Baharvand H

doi

10.1089/scd.2010.0517

subject

Has Abstract

pub_date

2011-11-01 00:00:00

pages

1911-23

issue

11

eissn

1547-3287

issn

1557-8534

journal_volume

20

pub_type

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