Bcl-xL enhances single-cell survival and expansion of human embryonic stem cells without affecting self-renewal

Hao Bai, Kang Chen, Yong Xing Gao, Melanie Arzigian, Yin Liang Xie, Christopher Malcosky, Yong Guang Yang, Wen Shu Wu, Zack Z. Wang

Research output: Contribution to journalArticle

Abstract

Robust expansion and genetic manipulation of human embryonic stem cells (hESCs) and induced-pluripotent stem (iPS) cells are limited by poor cell survival after enzymatic dissociation into single cells. Although inhibition of apoptosis is implicated for the single-cell survival of hESCs, the protective role of attenuation of apoptosis in hESC survival has not been elucidated. Bcl-xL is one of several anti-apoptotic proteins, which are members of the Bcl-2 family of proteins. Using an inducible system, we ectopically expressed Bcl-xL gene in hESCs, and found a significant increase of hESC colonies in the single-cell suspension cultures. Overexpression of Bcl-xL in hESCs decreased apoptotic caspase-3 + cells, suggesting attenuation of apoptosis in hESCs. Without altering the kinetics of pluripotent gene expression, the efficiency to generate embryoid bodies (EBs) in vitro and the formation of teratoma in vivo were significantly increased in Bcl-xL-overexpressing hESCs after single-cell dissociation. Interestingly, the number and size of hESC colonies from cluster cultures were not affected by Bcl-xL overexpression. Several genes of extracellular matrix and adhesion molecules were upregulated by Bcl-xL in hESCs without single-cell dissociation, suggesting that Bcl-xL regulates adhesion molecular expression independent of cell dissociation. In addition, the gene expressions of FAS and several TNF signaling mediators were downregulated by Bcl-xL.These data support a model in which Bcl-xL promotes cell survival and increases cloning efficiency of dissociated hESCs without altering hESC self-renewal by i) attenuation of apoptosis, and ii) upregulation of adhesion molecules to facilitate cell-cell or cell-matrix interactions.

Original languageEnglish (US)
Pages (from-to)26-37
Number of pages12
JournalStem Cell Research
Volume8
Issue number1
DOIs
StatePublished - Jan 2012
Externally publishedYes

Fingerprint

Cell Survival
Apoptosis
Human Embryonic Stem Cells
Embryoid Bodies
Gene Expression
Induced Pluripotent Stem Cells
Apoptosis Regulatory Proteins
Teratoma
Cell Communication
Caspase 3
Genes
Extracellular Matrix
Organism Cloning
Suspensions
Up-Regulation
Down-Regulation
Cell Culture Techniques

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Medicine(all)

Cite this

Bcl-xL enhances single-cell survival and expansion of human embryonic stem cells without affecting self-renewal. / Bai, Hao; Chen, Kang; Gao, Yong Xing; Arzigian, Melanie; Xie, Yin Liang; Malcosky, Christopher; Yang, Yong Guang; Wu, Wen Shu; Wang, Zack Z.

In: Stem Cell Research, Vol. 8, No. 1, 01.2012, p. 26-37.

Research output: Contribution to journalArticle

Bai, Hao ; Chen, Kang ; Gao, Yong Xing ; Arzigian, Melanie ; Xie, Yin Liang ; Malcosky, Christopher ; Yang, Yong Guang ; Wu, Wen Shu ; Wang, Zack Z. / Bcl-xL enhances single-cell survival and expansion of human embryonic stem cells without affecting self-renewal. In: Stem Cell Research. 2012 ; Vol. 8, No. 1. pp. 26-37.
@article{f0a544b22b944916989f9e203cdd9c96,
title = "Bcl-xL enhances single-cell survival and expansion of human embryonic stem cells without affecting self-renewal",
abstract = "Robust expansion and genetic manipulation of human embryonic stem cells (hESCs) and induced-pluripotent stem (iPS) cells are limited by poor cell survival after enzymatic dissociation into single cells. Although inhibition of apoptosis is implicated for the single-cell survival of hESCs, the protective role of attenuation of apoptosis in hESC survival has not been elucidated. Bcl-xL is one of several anti-apoptotic proteins, which are members of the Bcl-2 family of proteins. Using an inducible system, we ectopically expressed Bcl-xL gene in hESCs, and found a significant increase of hESC colonies in the single-cell suspension cultures. Overexpression of Bcl-xL in hESCs decreased apoptotic caspase-3 + cells, suggesting attenuation of apoptosis in hESCs. Without altering the kinetics of pluripotent gene expression, the efficiency to generate embryoid bodies (EBs) in vitro and the formation of teratoma in vivo were significantly increased in Bcl-xL-overexpressing hESCs after single-cell dissociation. Interestingly, the number and size of hESC colonies from cluster cultures were not affected by Bcl-xL overexpression. Several genes of extracellular matrix and adhesion molecules were upregulated by Bcl-xL in hESCs without single-cell dissociation, suggesting that Bcl-xL regulates adhesion molecular expression independent of cell dissociation. In addition, the gene expressions of FAS and several TNF signaling mediators were downregulated by Bcl-xL.These data support a model in which Bcl-xL promotes cell survival and increases cloning efficiency of dissociated hESCs without altering hESC self-renewal by i) attenuation of apoptosis, and ii) upregulation of adhesion molecules to facilitate cell-cell or cell-matrix interactions.",
author = "Hao Bai and Kang Chen and Gao, {Yong Xing} and Melanie Arzigian and Xie, {Yin Liang} and Christopher Malcosky and Yang, {Yong Guang} and Wu, {Wen Shu} and Wang, {Zack Z.}",
year = "2012",
month = "1",
doi = "10.1016/j.scr.2011.08.002",
language = "English (US)",
volume = "8",
pages = "26--37",
journal = "Stem Cell Research",
issn = "1873-5061",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Bcl-xL enhances single-cell survival and expansion of human embryonic stem cells without affecting self-renewal

AU - Bai, Hao

AU - Chen, Kang

AU - Gao, Yong Xing

AU - Arzigian, Melanie

AU - Xie, Yin Liang

AU - Malcosky, Christopher

AU - Yang, Yong Guang

AU - Wu, Wen Shu

AU - Wang, Zack Z.

PY - 2012/1

Y1 - 2012/1

N2 - Robust expansion and genetic manipulation of human embryonic stem cells (hESCs) and induced-pluripotent stem (iPS) cells are limited by poor cell survival after enzymatic dissociation into single cells. Although inhibition of apoptosis is implicated for the single-cell survival of hESCs, the protective role of attenuation of apoptosis in hESC survival has not been elucidated. Bcl-xL is one of several anti-apoptotic proteins, which are members of the Bcl-2 family of proteins. Using an inducible system, we ectopically expressed Bcl-xL gene in hESCs, and found a significant increase of hESC colonies in the single-cell suspension cultures. Overexpression of Bcl-xL in hESCs decreased apoptotic caspase-3 + cells, suggesting attenuation of apoptosis in hESCs. Without altering the kinetics of pluripotent gene expression, the efficiency to generate embryoid bodies (EBs) in vitro and the formation of teratoma in vivo were significantly increased in Bcl-xL-overexpressing hESCs after single-cell dissociation. Interestingly, the number and size of hESC colonies from cluster cultures were not affected by Bcl-xL overexpression. Several genes of extracellular matrix and adhesion molecules were upregulated by Bcl-xL in hESCs without single-cell dissociation, suggesting that Bcl-xL regulates adhesion molecular expression independent of cell dissociation. In addition, the gene expressions of FAS and several TNF signaling mediators were downregulated by Bcl-xL.These data support a model in which Bcl-xL promotes cell survival and increases cloning efficiency of dissociated hESCs without altering hESC self-renewal by i) attenuation of apoptosis, and ii) upregulation of adhesion molecules to facilitate cell-cell or cell-matrix interactions.

AB - Robust expansion and genetic manipulation of human embryonic stem cells (hESCs) and induced-pluripotent stem (iPS) cells are limited by poor cell survival after enzymatic dissociation into single cells. Although inhibition of apoptosis is implicated for the single-cell survival of hESCs, the protective role of attenuation of apoptosis in hESC survival has not been elucidated. Bcl-xL is one of several anti-apoptotic proteins, which are members of the Bcl-2 family of proteins. Using an inducible system, we ectopically expressed Bcl-xL gene in hESCs, and found a significant increase of hESC colonies in the single-cell suspension cultures. Overexpression of Bcl-xL in hESCs decreased apoptotic caspase-3 + cells, suggesting attenuation of apoptosis in hESCs. Without altering the kinetics of pluripotent gene expression, the efficiency to generate embryoid bodies (EBs) in vitro and the formation of teratoma in vivo were significantly increased in Bcl-xL-overexpressing hESCs after single-cell dissociation. Interestingly, the number and size of hESC colonies from cluster cultures were not affected by Bcl-xL overexpression. Several genes of extracellular matrix and adhesion molecules were upregulated by Bcl-xL in hESCs without single-cell dissociation, suggesting that Bcl-xL regulates adhesion molecular expression independent of cell dissociation. In addition, the gene expressions of FAS and several TNF signaling mediators were downregulated by Bcl-xL.These data support a model in which Bcl-xL promotes cell survival and increases cloning efficiency of dissociated hESCs without altering hESC self-renewal by i) attenuation of apoptosis, and ii) upregulation of adhesion molecules to facilitate cell-cell or cell-matrix interactions.

UR - http://www.scopus.com/inward/record.url?scp=80052592682&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80052592682&partnerID=8YFLogxK

U2 - 10.1016/j.scr.2011.08.002

DO - 10.1016/j.scr.2011.08.002

M3 - Article

C2 - 22099018

AN - SCOPUS:80052592682

VL - 8

SP - 26

EP - 37

JO - Stem Cell Research

JF - Stem Cell Research

SN - 1873-5061

IS - 1

ER -