Research ArticleVascular biology Open Access | 10.1172/jci.insight.164781

Yang Lin,1,2 Kimihiko Banno,1,3 Chang-Hyun Gil,1,4 Jered Myslinski,5 Takashi Hato,5 William C. Shelley,1,4 Hongyu Gao,6 Xiaoling Xuei,6 Yunlong Liu,6 David P. Basile,7 Momoko Yoshimoto,1,8 Nutan Prasain,1 Stefan P. Tarnawsky,1 Ralf H. Adams,9 Katsuhiko Naruse,10 Junko Yoshida,3 Michael P. Murphy,4 Kyoji Horie,3 and Mervin C. Yoder1,4,11

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Lin, Y. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Banno, K. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Gil, C. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Myslinski, J. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Hato, T. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Shelley, W. in: JCI | PubMed | Google Scholar

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Gao, H. in: JCI | PubMed | Google Scholar

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Xuei, X. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Liu, Y. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Basile, D. in: JCI | PubMed | Google Scholar

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Yoshimoto, M. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Prasain, N. in: JCI | PubMed | Google Scholar

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Tarnawsky, S. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Adams, R. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Naruse, K. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Yoshida, J. in: JCI | PubMed | Google Scholar

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Murphy, M. in: JCI | PubMed | Google Scholar

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Horie, K. in: JCI | PubMed | Google Scholar |

1Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.

2Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

3Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan.

4Department of Surgery,

5Department of Medicine,

6Department of Medical and Molecular Genetics, and

7Department of Anatomy Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, Indiana, USA.

8Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, USA.

9Max Planck Institute for Molecular Biomedicine, Muenster, Germany.

10Department of Obstetrics & Gynecology, Nara Medical University, Kashihara, Nara, Japan.

11Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Address correspondence to: Kimihiko Banno, Department of Physiology II, Nara Medical University, 840 Shijo-sho, Kashihara, Nara, Japan. Phone 81.744.29.8829; Email: kbanno@naramed-u.ac.jp. Or to: Mervin C. Yoder, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine, 975 W. Walnut St., Indianapolis, Indiana, USA. Phone 317.274.9787; Email: myoder@iu.edu.

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Find articles by Yoder, M. in: JCI | PubMed | Google Scholar |

Authorship note: YL, KB, and CHG contributed equally to this work. KB and MCY are co–corresponding authors.

Published January 24, 2023 - More info

Published in Volume 8, Issue 5 on March 8, 2023
JCI Insight. 2023;8(5):e164781. https://doi.org/10.1172/jci.insight.164781.
© 2023 Lin et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Published January 24, 2023 - Version history
Received: August 24, 2022; Accepted: January 18, 2023 View PDF Abstract

Most circulating endothelial cells are apoptotic, but rare circulating endothelial colony-forming cells (C-ECFCs), also known as blood outgrowth endothelial cells, with proliferative and vasculogenic activity can be cultured; however, the origin and naive function of these C-ECFCs remains obscure. Herein, detailed lineage tracing revealed murine C-ECFCs emerged in the early postnatal period, displayed high vasculogenic potential with enriched frequency of clonal proliferative cells compared with tissue-resident ECFCs, and were not committed to or derived from the BM hematopoietic system but from tissue-resident ECFCs. In humans, C-ECFCs were present in the CD34bright cord blood mononuclear subset, possessed proliferative potential and in vivo vasculogenic function in a naive or cultured state, and displayed a single cell transcriptome sharing some umbilical venous endothelial cell features, such as a higher protein C receptor and extracellular matrix gene expression. This study provides an advance for the field by identifying the origin, naive function, and antigens to prospectively isolate C-ECFCs for translational studies.

Graphical Abstract Introduction

Circulating endothelial cells (C-ECs) are dead or dying ECs sloughed off tissue vasculature (1, 2), and their number serves as a biomarker for various disease states (3, 4) (Table 1, “List of nonstandard abbreviations”). In humans, circulating endothelial colony-forming cells (C-ECFCs), also known as blood outgrowth endothelial cells, with high clonal proliferative potential are a subset within C-ECs in umbilical cord blood (CB) or adult blood (5–8). C-ECFCs are vasculogenic cells displaying in vivo vessel-forming ability, discriminating their functions from BM-derived endothelial progenitor cells (EPCs) (9) restricted to playing proangiogenic roles (10–12). However, several important unaddressed questions remain: Where do C-ECFCs originate, and do C-ECFCs in their in vivo naive state function similarly to isolated, growth factor–expanded, and cultured ECFCs?

Table 1

List of nonstandard abbreviations in this manuscript

We have addressed these questions in mice and found murine C-ECFCs in a developmentally regulated window across several mouse strains. We identified lineage-traced C-ECFCs with high proliferative potential that upon transplantation in a naive or cultured expanded state form robust vasculature. Moreover, single-cell RNA-Seq analysis (scRNA-Seq) of CB distinguished various C-EC subsets. Markers defining the subsets permitted prospective isolation of C-ECFCs from CB with evidence that naive or cultured C-ECFCs display clonal proliferative potential and in vivo vessel formation, defining these cells as a distinct subpopulation of C-ECs. We have thus uncovered key characteristics of C-ECFCs in mice and humans, with information to accelerate C-ECFCs as a viable choice for cell therapy.

Results

C-ECFCs in mice possess vasculogenic potential similar to humans. C-ECFCs, though present in humans, are not demonstrable in individual adult mice. Previously, blood from 5–10 mice (>8-week-old C57BL/6J animals) was pooled into a single aliquot and plated in culture and in 5 of 44 (11%) attempts (total 282 mice) was reportedly successful in growing at least 1 C-ECFC colony (13). Since CB ECFCs are enriched 60- to 100-fold compared with adult peripheral blood (PB) (6), we isolated murine PBMCs from prenatal animals to 3 months of age and seeded mononuclear cells (MCs) on OP9 feeder cells (Supplemental Figure 1A; supplemental material available online with this article;