Nano-structural analysis of engrafted human induced pluripotent stem cell-derived cardiomyocytes in mouse hearts using a genetic-probe APEX2

Takeshi Hatani; Shunsuke Funakoshi; Thomas J Deerinck; Eric A Bushong; Takeshi Kimura; Hiroshi Takeshima; Mark H Ellisman; Masahiko Hoshijima; Yoshinori Yoshida

doi:10.1016/j.bbrc.2018.10.020

Nano-structural analysis of engrafted human induced pluripotent stem cell-derived cardiomyocytes in mouse hearts using a genetic-probe APEX2

Biochem Biophys Res Commun. 2018 Nov 10;505(4):1251-1256. doi: 10.1016/j.bbrc.2018.10.020. Epub 2018 Oct 15.

Authors

Takeshi Hatani¹, Shunsuke Funakoshi², Thomas J Deerinck³, Eric A Bushong³, Takeshi Kimura⁴, Hiroshi Takeshima⁵, Mark H Ellisman⁶, Masahiko Hoshijima⁷, Yoshinori Yoshida⁸

Affiliations

¹ Department of Cardiovascular Medicine, Graduate School of Medicine, Kyoto University, Kyoto, 606-8507, Japan; Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
² Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
³ Center for Research in Biological Systems, University of California at San Diego, La Jolla, CA, 92093, USA; National Center for Microscopy and Imaging Research, University of California at San Diego, La Jolla, CA, 92093, USA.
⁴ Department of Cardiovascular Medicine, Graduate School of Medicine, Kyoto University, Kyoto, 606-8507, Japan.
⁵ Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, 606-8501, Japan.
⁶ Center for Research in Biological Systems, University of California at San Diego, La Jolla, CA, 92093, USA; National Center for Microscopy and Imaging Research, University of California at San Diego, La Jolla, CA, 92093, USA; Departments of Neurosciences and Bioengineering, University of California at San Diego, La Jolla, CA, 92093, USA.
⁷ Center for Research in Biological Systems, University of California at San Diego, La Jolla, CA, 92093, USA; Department of Medicine, University of California at San Diego, La Jolla, CA, 92093, USA. Electronic address: [email protected].
⁸ Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan. Electronic address: [email protected].

Abstract

Many studies have shown the feasibility of in vivo cardiac transplantation of human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) in animal experiments. However, nano-structural confirmation of the successful incorporation of the engrafted iPSC-CMs including electron microscopy (EM) has not been accomplished, partly because identification of graft cells in EM has proven to be difficult. Using APEX2, an engineered ascorbate peroxidase imaging tag, we successfully localized and analyzed the fine structure of sarcomeres and the excitation contraction machinery of iPSC-CMs 6 months after their engraftment in infarcted mouse hearts. APEX2 made iPSC-CMs visible in multiple imaging modalities including light microscopy, X-ray microscopic tomography, transmission EM, and scanning EM. EM tomography allowed assessment of the differentiation state of APEX2-positive iPSC-CMs and analysis of the fine structure of the sarcomeres including T-tubules and dyads.

Keywords: APEX2; Cardiac cell transplantation; Electron microscopy; Regeneration therapy; iPSC-derived cardiomyocytes.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation
Cell Line
Cell Lineage
DNA-(Apurinic or Apyrimidinic Site) Lyase / genetics
Heart / anatomy & histology
Humans
Induced Pluripotent Stem Cells / cytology*
Male
Mice
Molecular Probes
Myocardial Infarction / pathology
Myocardium / cytology*
Myocardium / ultrastructure
Myocytes, Cardiac / cytology
Myocytes, Cardiac / transplantation*

Substances

Molecular Probes
DNA-(Apurinic or Apyrimidinic Site) Lyase

Abstract

Publication types

MeSH terms

Substances

Grants and funding