Endothelial properties of third-trimester amniotic fluid stem cells cultured in hypoxia

Stem Cell Research & Therapy

Michela Pozzobon¹  Paolo De Coppi⁴  Gian Paolo Fadini²  Erich Cosmi³  Silvia Visentin³  Luca Urbani⁴  Enrica Bertin¹  Giovanna Spiro²  Martina Piccoli¹  Mattia Albiero²  Chiara Franzin¹  Andrea Alex Schiavo³ 

[1] Stem Cells and Regenerative Medicine Laboratory, Foundation Institute of Pediatric Research Città della Speranza, Corso Stati Uniti 4, Padova, 35127, Italy;Medicine Department (DIMED), University of Padova, via Giustiniani 2, Padova, 35100, Italy;Department of Woman and Children Health, University of Padova, via Giustinani 2, Padova, 35100, Italy;Stem Cells and Regenerative Medicine Section, Developmental biology and Cancer Program, Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1EH, UK

关键词: Ischemia;    Endothelial dysfunction;    Tissue regeneration;    Fetal stem cells;    c-Kit;   

Others  :  1234416
DOI  :  10.1186/s13287-015-0204-0

 received in 2015-06-24, accepted in 2015-10-15,  发布年份 2015

【 摘 要 】

Introduction

Endothelial dysfunction is found in different pathologies such as diabetes and renal and heart diseases, representing one of the major health problems. The reduced vasodilation of impaired endothelium starts a prothrombotic state associated with irregular blood flow. We aimed to explore the potential of amniotic fluid stem (AFS) cells as a source for regenerative medicine in this field; for the first time, we focused on third-trimester amniotic fluid AFS cells and compared them with the already-described AFS cells from the second trimester.

Methods

Cells from the two trimesters were cultured, selected and expanded in normoxia (20 % oxygen) and hypoxia (5 % oxygen). Cells were analysed to compare markers, proliferation rate and differentiation abilities. Endothelial potential was assessed not only in vitro—Matrigel tube formation assay, acetylated human low-density lipoprotein (AcLDL) uptake—but also in vivo (Matrigel plug with cell injection and two animal models). Specifically, for the latter, we used established protocols to assess the involvement of AFS cells in two different mouse models of endothelial dysfunction: (1) a chronic ischemia model with local injection of cells and (2) an electric carotid damage where cells were systemically injected.

Results

We isolated and expanded AFS cells from third-trimester amniotic fluid samples by using CD117 as a selection marker. Hypoxia enhanced the proliferation rate, the surface protein pattern was conserved between the trimesters and comparable differentiation was achieved after culture in both normoxia and hypoxia. Notably, the expression of early endothelial transcription factors and AngiomiRs was detected before and after induction. When in vivo, AFS cells from both trimesters expanded in hypoxia were able to rescue the surface blood flow when locally injected in mice after chronic ischemia damage, and importantly AFS cells at term of gestation possessed enhanced ability to fix carotid artery electric damage compared with AFS cells from the second trimester.

Conclusions

To the best of our knowledge, this is the first research work that fully characterizes AFS cells from the third trimester for regenerative medicine purposes. The results highlight how AFS cells, in particular at term of gestation and cultured in hypoxia, can be considered a promising source of stem cells possessing significant endothelial regenerative potential.

【 授权许可】

   
2015 Schiavo et al.

【 图 表 】

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