A protocol for transdifferentiation of human cardiac fibroblasts into endothelial cells via activation of innate immunity

Chun Liu; Pedro Medina; Dilip Thomas; Ian Y Chen; Karim Sallam; Danish Sayed; Nazish Sayed

doi:10.1016/j.xpro.2021.100556

A protocol for transdifferentiation of human cardiac fibroblasts into endothelial cells via activation of innate immunity

STAR Protoc. 2021 Jun 5;2(2):100556. doi: 10.1016/j.xpro.2021.100556. eCollection 2021 Jun 18.

Authors

Chun Liu^{1

2}, Pedro Medina¹, Dilip Thomas^{1

2}, Ian Y Chen^{1

2}, Karim Sallam^{1

2}, Danish Sayed³, Nazish Sayed^{1

2

4}

Affiliations

¹ Stanford Cardiovascular Institute, Stanford University, Stanford, CA 94305, USA.
² Department of Medicine, Stanford University, Stanford, CA 94305, USA.
³ Department of Cell Biology and Molecular Medicine, Rutgers New Jersey Medical School, Newark, NJ 07103, USA.
⁴ Department of Surgery, Division of Vascular Surgery, Stanford University, Stanford, CA 94305, USA.

Abstract

Endothelial cells (ECs) have emerged as key pathogenic players in cardiac disease due to their proximity with cardiomyocytes. Induced pluripotent stem cells (iPSCs) have been employed to generate ECs. However, it may be more clinically relevant to transdifferentiate fibroblasts into ECs directly without introducing pluripotent or virally driven transcription factors. Here, we present a protocol that describes the direct conversion of human cardiac fibroblasts into ECs by leveraging the innate immune system. Our protocol produces bona fide human ECs with 95%-98% purity by first passage. For complete details on the use and execution of this protocol, please refer to Liu et al. (2020) and Sayed et al. (2015).

Keywords: Cell Differentiation; Cell culture; Flow Cytometry/Mass Cytometry; Molecular Biology; Stem Cells.

Publication types

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

MeSH terms

Cell Transdifferentiation*
Culture Media
Endothelial Cells / cytology
Fibroblasts / cytology
Humans
Immunity, Innate*
Myocytes, Cardiac / cytology*
Transcription Factors / metabolism

Substances

Culture Media
Transcription Factors

Grants and funding

K01 HL135455/HL/NHLBI NIH HHS/United States