Hypoxic Preconditioning Enhances Survival and Proangiogenic Capacity of Human First Trimester Chorionic Villus-Derived Mesenchymal Stem Cells for Fetal Tissue Engineering.

Dake Hao,Lee Lankford,Diana L Farmer,Bowen Ma,Lizette Reynaga,Fuzheng Guo,Aijun Wang,Chuanchao He

doi:10.1155/2019/9695239

Abstract

Prenatal stem cell-based regenerative therapies have progressed substantially and have been demonstrated as effective treatment options for fetal diseases that were previously deemed untreatable. Due to immunoregulatory properties, self-renewal capacity, and multilineage potential, autologous human placental chorionic villus-derived mesenchymal stromal cells (CV-MSCs) are an attractive cell source for fetal regenerative therapies. However, as a general issue for MSC transplantation, the poor survival and engraftment is a major challenge of the application of MSCs. Particularly for the fetal transplantation of CV-MSCs in the naturally hypoxic fetal environment, improving the survival and engraftment of CV-MSCs is critically important. Hypoxic preconditioning (HP) is an effective priming approach to protect stem cells from ischemic damage. In this study, we developed an optimal HP protocol to enhance the survival and proangiogenic capacity of CV-MSCs for improving clinical outcomes in fetal applications. Total cell number, DNA quantification, nuclear area test, and cell viability test showed HP significantly protected CV-MSCs from ischemic damage. Flow cytometry analysis confirmed HP did not alter the immunophenotype of CV-MSCs. Caspase-3, MTS, and Western blot analysis showed HP significantly reduced the apoptosis of CV-MSCs under ischemic stimulus via the activation of the AKT signaling pathway that was related to cell survival. ELISA results showed HP significantly enhanced the secretion of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) by CV-MSCs under an ischemic stimulus. We also found that the environmental nutrition level was critical for the release of brain-derived neurotrophic factor (BDNF). The angiogenesis assay results showed HP-primed CV-MSCs could significantly enhance endothelial cell (EC) proliferation, migration, and tube formation. Consequently, HP is a promising strategy to increase the tolerance of CV-MSCs to ischemia and improve their therapeutic efficacy in fetal clinical applications.

Highlights

Over the past three decades, with the development and exciting advances of fetal surgery for treatment of congenital diseases, fetal tissue engineering has been established as an emerging field of fetal medicine to augment in utero surgical approaches [1,2,3]
For the effect of the hypoxic condition on the growth factor release of chorionic villus-derived mesenchymal stromal cells (CV-Mesenchymal stem cells (MSCs)), the results showed that the release of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) from CV-MSCs cultured in D5 media were significantly higher in the hypoxic condition compared to the normoxic condition and that there was no significant difference for brain-derived neurotrophic factor (BDNF) release (Figure 6(a))
We evaluated the effect of Hypoxic preconditioning (HP) on CV-MSC secretion and demonstrated that secretion of several cytokines related to vascular protection, such as VEGF and HGF, was significantly higher in the HP group, compared to the non-HP group under ischemic stimulus

Summary

Introduction

Over the past three decades, with the development and exciting advances of fetal surgery for treatment of congenital diseases, fetal tissue engineering has been established as an emerging field of fetal medicine to augment in utero surgical approaches [1,2,3]. Mesenchymal stem cells (MSCs) are multipotent stem cells with the ability to self-renew and have been isolated from various tissues, such as the bone marrow [4], heart [5], adipose [6], peripheral blood [7], dental pulp [8], cord blood [9], menstrual blood [10,11,12], Wharton’ s jelly [13], and chorionic villi [14]. Isolation of MSCs from first trimester chorionic villus tissue (CV-MSCs) that allows for therapeutic use for in utero applications represents a promising approach for autologous fetal treatment of birth defects [38]

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Conclusion