Comparison of Anti-Oxidative Effect of Human Adipose- and Amniotic Membrane-Derived Mesenchymal Stem Cell Conditioned Medium on Mouse Preimplantation Embryo Development.

Kihae Ra,Sung Keun Kang,Hyun Ju Oh,Byeong Chun Lee,Eui Hyun Kim,Eun Young Kim,Se Chang Park,Jeong Chan Ra

doi:10.3390/antiox10020268

Abstract

Oxidative stress is a major cause of damage to the quantity and quality of embryos produced in vitro. Antioxidants are usually supplemented to protect embryos from the suboptimal in vitro culture (IVC) environment. Amniotic membrane-derived mesenchymal stem cells (AMSC) have emerged as a promising regenerative therapy, and their paracrine factors with anti-oxidative effects are present in AMSC conditioned medium (CM). We examined the anti-oxidative potential of human AMSC-CM treatment during IVC on mouse preimplantation embryo development and antioxidant gene expression in the forkhead box O (FoxO) pathway. AMSC-CM (10%) was optimal for overall preimplantation embryo developmental processes and upregulated the expression of FoxOs and their downstream antioxidants in blastocysts (BL). Subsequently, compared to adipose-derived mesenchymal stem cell (ASC)-CM, AMSC-CM enhanced antioxidant gene expression and intracellular GSH levels in the BL. Total antioxidant capacity and SOD activity were greater in AMSC-CM than in ASC-CM. Furthermore, SOD and catalase were more active in culture medium supplemented with AMSC-CM than in ASC-CM. Lastly, the anti-apoptotic effect of AMSC-CM was observed with the regulation of apoptosis-related genes and mitochondrial membrane potential in BL. In conclusion, the present study established AMSC-CM treatment at an optimal concentration as a novel antioxidant intervention for assisted reproduction.

Highlights

Amniotic membrane-derived mesenchymal stem cells (AMSC) were analyzed with flow cytometry to identify the expression of phenotypic markers (Figure 1) and confirmed that AMSCs from all donors were positive for mesenchymal markers (CD73, CD90, CD105, CD29, and CD44), and negative for the endothelial marker (CD31) and hematopoietic markers (CD34 and CD45)
Embryo development to 4, 16-cell stages, BL, and hatched BL was evaluated to determine the optimal concentration of AMSC-conditioned medium (CM) supplementation among the three different concentrations of AMSC-CM (10%, 20%, and 50%)
This study established that AMSC-CM treatment, at the optimal concentration, acts as an antioxidant during in vitro culture (IVC) of mouse preimplantation embryos

Summary

Introduction

The success rate of assisted reproductive technologies (ART) to surmount infertility has increased with the improvement of conditions for embryo in vitro production [1]. The balance of reactive oxygen species (ROS) and antioxidants is maintained at physiologically normal levels in female reproductive systems, but is disrupted in vitro, resulting in an increase in exposure to oxidative damage risk [2]. In the process of assisted reproduction, a number of external factors causing oxidative stress appear from technical procedures to environmental sources [3]. Oxidative stress due to accumulated ROS in in vitro-produced embryos impairs the efficiency of embryonic development and induces reproductive failure due to an increase in embryo fragmentation and apoptosis, and a Antioxidants 2021, 10, 268.

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