Microfluidic single-cell analysis of transplanted human induced pluripotent stem cell-derived cardiomyocytes after acute myocardial infarction

Sang Ging Ong, Bruno C. Huber, Won Hee Lee, Kazuki Kodo, Antje D. Ebert, Yu Ma, Patricia K. Nguyen, Sebastian Diecke, Wen Yi Chen, Joseph C. Wu

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Background - Human induced pluripotent stem cells (iPSCs) are attractive candidates for therapeutic use, with the potential to replace deficient cells and to improve functional recovery in injury or disease settings. Here, we test the hypothesis that human iPSC-derived cardiomyocytes (iPSC-CMs) can secrete cytokines as a molecular basis to attenuate adverse cardiac remodeling after myocardial infarction. Methods and Results - Human iPSCs were generated from skin fibroblasts and differentiated in vitro with a small molecule-based protocol. Troponin+ iPSC-CMs were confirmed by immunohistochemistry, quantitative polymerase chain reaction, fluorescence-activated cell sorting, and electrophysiological measurements. Afterward, 2×106 iPSC-CMs derived from a cell line transduced with a vector expressing firefly luciferase and green fluorescent protein were transplanted into adult NOD/SCID mice with acute left anterior descending artery ligation. Control animals received PBS injection. Bioluminescence imaging showed limited engraftment on transplantation into ischemic myocardium. However, magnetic resonance imaging of animals transplanted with iPSC-CMs showed significant functional improvement and attenuated cardiac remodeling compared with PBS-treated control animals. To understand the underlying molecular mechanism, microfluidic single-cell profiling of harvested iPSC-CMs, laser capture microdissection of host myocardium, and in vitro ischemia stimulation were used to demonstrate that the iPSC-CMs could release significant levels of proangiogenic and antiapoptotic factors in the ischemic microenvironment. Conclusions - Transplantation of human iPSC-CMs into an acute mouse myocardial infarction model can improve left ventricular function and attenuate cardiac remodeling. Because of limited engraftment, most of the effects are possibly explained by paracrine activity of these cells.

Original languageEnglish
Pages (from-to)762-771
Number of pages10
JournalCirculation
Volume132
Issue number8
DOIs
Publication statusPublished - 2015 Aug 25
Externally publishedYes

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Single-Cell Analysis
Induced Pluripotent Stem Cells
Microfluidics
Cardiac Myocytes
Myocardial Infarction
Myocardium
Transplantation
Laser Capture Microdissection
Firefly Luciferases
Inbred NOD Mouse
Troponin
SCID Mice
Therapeutic Uses
Green Fluorescent Proteins
Left Ventricular Function
Ligation
Flow Cytometry
Ischemia
Arteries
Fibroblasts

Keywords

  • cell transplantation
  • molecular imaging
  • myocardial infarction
  • myocytes cardiac
  • paracrine communication
  • stem cells

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Microfluidic single-cell analysis of transplanted human induced pluripotent stem cell-derived cardiomyocytes after acute myocardial infarction. / Ong, Sang Ging; Huber, Bruno C.; Lee, Won Hee; Kodo, Kazuki; Ebert, Antje D.; Ma, Yu; Nguyen, Patricia K.; Diecke, Sebastian; Chen, Wen Yi; Wu, Joseph C.

In: Circulation, Vol. 132, No. 8, 25.08.2015, p. 762-771.

Research output: Contribution to journalArticle

Ong, Sang Ging ; Huber, Bruno C. ; Lee, Won Hee ; Kodo, Kazuki ; Ebert, Antje D. ; Ma, Yu ; Nguyen, Patricia K. ; Diecke, Sebastian ; Chen, Wen Yi ; Wu, Joseph C. / Microfluidic single-cell analysis of transplanted human induced pluripotent stem cell-derived cardiomyocytes after acute myocardial infarction. In: Circulation. 2015 ; Vol. 132, No. 8. pp. 762-771.
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AU - Huber, Bruno C.

AU - Lee, Won Hee

AU - Kodo, Kazuki

AU - Ebert, Antje D.

AU - Ma, Yu

AU - Nguyen, Patricia K.

AU - Diecke, Sebastian

AU - Chen, Wen Yi

AU - Wu, Joseph C.

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N2 - Background - Human induced pluripotent stem cells (iPSCs) are attractive candidates for therapeutic use, with the potential to replace deficient cells and to improve functional recovery in injury or disease settings. Here, we test the hypothesis that human iPSC-derived cardiomyocytes (iPSC-CMs) can secrete cytokines as a molecular basis to attenuate adverse cardiac remodeling after myocardial infarction. Methods and Results - Human iPSCs were generated from skin fibroblasts and differentiated in vitro with a small molecule-based protocol. Troponin+ iPSC-CMs were confirmed by immunohistochemistry, quantitative polymerase chain reaction, fluorescence-activated cell sorting, and electrophysiological measurements. Afterward, 2×106 iPSC-CMs derived from a cell line transduced with a vector expressing firefly luciferase and green fluorescent protein were transplanted into adult NOD/SCID mice with acute left anterior descending artery ligation. Control animals received PBS injection. Bioluminescence imaging showed limited engraftment on transplantation into ischemic myocardium. However, magnetic resonance imaging of animals transplanted with iPSC-CMs showed significant functional improvement and attenuated cardiac remodeling compared with PBS-treated control animals. To understand the underlying molecular mechanism, microfluidic single-cell profiling of harvested iPSC-CMs, laser capture microdissection of host myocardium, and in vitro ischemia stimulation were used to demonstrate that the iPSC-CMs could release significant levels of proangiogenic and antiapoptotic factors in the ischemic microenvironment. Conclusions - Transplantation of human iPSC-CMs into an acute mouse myocardial infarction model can improve left ventricular function and attenuate cardiac remodeling. Because of limited engraftment, most of the effects are possibly explained by paracrine activity of these cells.

AB - Background - Human induced pluripotent stem cells (iPSCs) are attractive candidates for therapeutic use, with the potential to replace deficient cells and to improve functional recovery in injury or disease settings. Here, we test the hypothesis that human iPSC-derived cardiomyocytes (iPSC-CMs) can secrete cytokines as a molecular basis to attenuate adverse cardiac remodeling after myocardial infarction. Methods and Results - Human iPSCs were generated from skin fibroblasts and differentiated in vitro with a small molecule-based protocol. Troponin+ iPSC-CMs were confirmed by immunohistochemistry, quantitative polymerase chain reaction, fluorescence-activated cell sorting, and electrophysiological measurements. Afterward, 2×106 iPSC-CMs derived from a cell line transduced with a vector expressing firefly luciferase and green fluorescent protein were transplanted into adult NOD/SCID mice with acute left anterior descending artery ligation. Control animals received PBS injection. Bioluminescence imaging showed limited engraftment on transplantation into ischemic myocardium. However, magnetic resonance imaging of animals transplanted with iPSC-CMs showed significant functional improvement and attenuated cardiac remodeling compared with PBS-treated control animals. To understand the underlying molecular mechanism, microfluidic single-cell profiling of harvested iPSC-CMs, laser capture microdissection of host myocardium, and in vitro ischemia stimulation were used to demonstrate that the iPSC-CMs could release significant levels of proangiogenic and antiapoptotic factors in the ischemic microenvironment. Conclusions - Transplantation of human iPSC-CMs into an acute mouse myocardial infarction model can improve left ventricular function and attenuate cardiac remodeling. Because of limited engraftment, most of the effects are possibly explained by paracrine activity of these cells.

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