title: Production of endothelial progenitor cells from skin fibroblasts by direct reprogramming for clinical usages.
abstract: Endothelial progenitor cells (EPCs) play an important role in angiogenesis. However, they exist in limited numbers in the human body. This study was aimed to produce EPCs, for autologous transplantation, using direct reprogramming of skin fibroblasts under GMP-compliant conditions. Fibroblasts were collected and cultured from the skin in DMEM/F12 medium supplemented with 5% activated platelet-rich plasma and 1% antibiotic-antimycotic solution. They were then transfected with mRNA ETV2 and incubated in culture medium under hypoxia (5% oxygen) for 14 d. Phenotype analysis of transfected cells confirmed that single-factor ETV2 transfection successfully reprogrammed dermal fibroblasts into functional EPCs. Our results showed that ETV2 mRNA combined with hypoxia can give rise to functional EPCs. The cells exhibited functional phenotypes similar to endothelial cells derived from umbilical cord vein; they expressed CD31 and VEGFR2, and formed capillary-like structures in vitro. Moreover, these EPCs could significantly improve hindlimb ischemia in mouse models. Although the direct conversion efficacy was low (3.12 ± 0.98%), altogether our study demonstrates that functional EPCs can be produced from fibroblasts and can be used in clinical applications.
authors: Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Ho Chi Minh City, Vietnam. pvphuc@hcmuns.edu.vn.
authors: Phuc
authors: P
authors: Van Pham
authors: Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Ho Chi Minh City, Vietnam.
authors: Ngoc Bich
authors: NB
authors: Vu
authors: Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Ho Chi Minh City, Vietnam.
authors: Thuy Thi-Thanh
authors: TT
authors: Dao
authors: Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Ho Chi Minh City, Vietnam.
authors: Ha Thi-Ngan
authors: HT
authors: Le
authors: Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Ho Chi Minh City, Vietnam.
authors: Lan Thi
authors: LT
authors: Phi
authors: Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Ho Chi Minh City, Vietnam.
authors: Ngoc Kim
authors: NK
authors: Phan
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D015687:Cell Hypoxia; D049109:Cell Proliferation / Q000235:genetics; D065150:Cellular Reprogramming / Q000235:genetics*; D066026:Endothelial Progenitor Cells / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology; D006614:Hindlimb / Q000166:cytology / Q000254:growth & development; D006801:Humans; D051379:Mice; D018919:Neovascularization, Physiologic / Q000235:genetics*; D012867:Skin / Q000166:cytology; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
chemical_list: C503994:ER71 protein, mouse; D014157:Transcription Factors
keywords: Direct conversion; ETV2; Endothelial progenitor cells; Epigenetic reprogramming; Hindlimb ischemia; Ischemia; Stem cell therapy