Differentiation of human induced-pluripotent stem cells into smooth-muscle cells: Two novel protocols

Libang Yang, Zhaohui Geng, Thomas Nickel, Caitlin Johnson, Lin Gao, James Dutton, Cody Hou, Jianyi Zhang

Research output: Contribution to journalArticle

  • 15 Citations

Abstract

Conventional protocols for differentiating human induced-pluripotent stem cells (hiPSCs) into smooth-muscle cells (SMCs) can be inefficient and generally fail to yield cells with a specific SMC phenotype (i.e., contractile or synthetic SMCs). Here, we present two novel hiPSC-SMC differentiation protocols that yield SMCs with predominantly contractile or synthetic phenotypes. Flow cytometry analyses of smooth-muscle actin (SMA) expression indicated that ∼45% of the cells obtained with each protocol assumed an SMC phenotype, and that the populations could be purified to ∼95% via metabolic selection. Assessments of cellular mRNA and/or protein levels indicated that SMA, myosin heavy chain II, collagen 1, calponin, transgelin, connexin 43, and vimentin expression in the SMCs obtained via the Contractile SMC protocol and in SMCs differentiated via a traditional protocol were similar, while SMCs produced via the Sythetic SMC protocol expressed less calponin, more collagen 1, and more connexin 43. Differences were also observed in functional assessments of the two SMC populations: the two-dimensional surface area of Contractile SMCs declined more extensively (to 12% versus 44% of original size) in response to carbachol treatment, while quantification of cell migration and proliferation were greater in Synthetic SMCs. Collectively, these data demonstrate that our novel differentiation protocols can efficiently generate SMCs from hiPSCs.

LanguageEnglish (US)
Article number0147155
JournalPLoS ONE
Volume11
Issue number1
DOIs
StatePublished - Jan 1 2016

Fingerprint

Induced Pluripotent Stem Cells
Stem cells
smooth muscle
myocytes
Smooth Muscle Myocytes
Muscle
Cells
Connexin 43
connexins
induced pluripotent stem cells
Phenotype
Actins
phenotype
Collagen
actin
collagen
Smooth Muscle Myosins
Functional assessment
Myosin Heavy Chains
carbachol

Cite this

Differentiation of human induced-pluripotent stem cells into smooth-muscle cells : Two novel protocols. / Yang, Libang; Geng, Zhaohui; Nickel, Thomas; Johnson, Caitlin; Gao, Lin; Dutton, James; Hou, Cody; Zhang, Jianyi.

In: PLoS ONE, Vol. 11, No. 1, 0147155, 01.01.2016.

Research output: Contribution to journalArticle

Yang, Libang ; Geng, Zhaohui ; Nickel, Thomas ; Johnson, Caitlin ; Gao, Lin ; Dutton, James ; Hou, Cody ; Zhang, Jianyi. / Differentiation of human induced-pluripotent stem cells into smooth-muscle cells : Two novel protocols. In: PLoS ONE. 2016 ; Vol. 11, No. 1.
@article{ca8f73f77324408a906485bdb243eb55,
title = "Differentiation of human induced-pluripotent stem cells into smooth-muscle cells: Two novel protocols",
abstract = "Conventional protocols for differentiating human induced-pluripotent stem cells (hiPSCs) into smooth-muscle cells (SMCs) can be inefficient and generally fail to yield cells with a specific SMC phenotype (i.e., contractile or synthetic SMCs). Here, we present two novel hiPSC-SMC differentiation protocols that yield SMCs with predominantly contractile or synthetic phenotypes. Flow cytometry analyses of smooth-muscle actin (SMA) expression indicated that ∼45{\%} of the cells obtained with each protocol assumed an SMC phenotype, and that the populations could be purified to ∼95{\%} via metabolic selection. Assessments of cellular mRNA and/or protein levels indicated that SMA, myosin heavy chain II, collagen 1, calponin, transgelin, connexin 43, and vimentin expression in the SMCs obtained via the Contractile SMC protocol and in SMCs differentiated via a traditional protocol were similar, while SMCs produced via the Sythetic SMC protocol expressed less calponin, more collagen 1, and more connexin 43. Differences were also observed in functional assessments of the two SMC populations: the two-dimensional surface area of Contractile SMCs declined more extensively (to 12{\%} versus 44{\%} of original size) in response to carbachol treatment, while quantification of cell migration and proliferation were greater in Synthetic SMCs. Collectively, these data demonstrate that our novel differentiation protocols can efficiently generate SMCs from hiPSCs.",
author = "Libang Yang and Zhaohui Geng and Thomas Nickel and Caitlin Johnson and Lin Gao and James Dutton and Cody Hou and Jianyi Zhang",
year = "2016",
month = "1",
day = "1",
doi = "10.1371/journal.pone.0147155",
language = "English (US)",
volume = "11",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "1",

}

TY - JOUR

T1 - Differentiation of human induced-pluripotent stem cells into smooth-muscle cells

T2 - PLoS One

AU - Yang,Libang

AU - Geng,Zhaohui

AU - Nickel,Thomas

AU - Johnson,Caitlin

AU - Gao,Lin

AU - Dutton,James

AU - Hou,Cody

AU - Zhang,Jianyi

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Conventional protocols for differentiating human induced-pluripotent stem cells (hiPSCs) into smooth-muscle cells (SMCs) can be inefficient and generally fail to yield cells with a specific SMC phenotype (i.e., contractile or synthetic SMCs). Here, we present two novel hiPSC-SMC differentiation protocols that yield SMCs with predominantly contractile or synthetic phenotypes. Flow cytometry analyses of smooth-muscle actin (SMA) expression indicated that ∼45% of the cells obtained with each protocol assumed an SMC phenotype, and that the populations could be purified to ∼95% via metabolic selection. Assessments of cellular mRNA and/or protein levels indicated that SMA, myosin heavy chain II, collagen 1, calponin, transgelin, connexin 43, and vimentin expression in the SMCs obtained via the Contractile SMC protocol and in SMCs differentiated via a traditional protocol were similar, while SMCs produced via the Sythetic SMC protocol expressed less calponin, more collagen 1, and more connexin 43. Differences were also observed in functional assessments of the two SMC populations: the two-dimensional surface area of Contractile SMCs declined more extensively (to 12% versus 44% of original size) in response to carbachol treatment, while quantification of cell migration and proliferation were greater in Synthetic SMCs. Collectively, these data demonstrate that our novel differentiation protocols can efficiently generate SMCs from hiPSCs.

AB - Conventional protocols for differentiating human induced-pluripotent stem cells (hiPSCs) into smooth-muscle cells (SMCs) can be inefficient and generally fail to yield cells with a specific SMC phenotype (i.e., contractile or synthetic SMCs). Here, we present two novel hiPSC-SMC differentiation protocols that yield SMCs with predominantly contractile or synthetic phenotypes. Flow cytometry analyses of smooth-muscle actin (SMA) expression indicated that ∼45% of the cells obtained with each protocol assumed an SMC phenotype, and that the populations could be purified to ∼95% via metabolic selection. Assessments of cellular mRNA and/or protein levels indicated that SMA, myosin heavy chain II, collagen 1, calponin, transgelin, connexin 43, and vimentin expression in the SMCs obtained via the Contractile SMC protocol and in SMCs differentiated via a traditional protocol were similar, while SMCs produced via the Sythetic SMC protocol expressed less calponin, more collagen 1, and more connexin 43. Differences were also observed in functional assessments of the two SMC populations: the two-dimensional surface area of Contractile SMCs declined more extensively (to 12% versus 44% of original size) in response to carbachol treatment, while quantification of cell migration and proliferation were greater in Synthetic SMCs. Collectively, these data demonstrate that our novel differentiation protocols can efficiently generate SMCs from hiPSCs.

UR - http://www.scopus.com/inward/record.url?scp=84955256208&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84955256208&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0147155

DO - 10.1371/journal.pone.0147155

M3 - Article

VL - 11

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 1

M1 - 0147155

ER -