Anti-Oxidative Effects of Human Adipose Stem Cell Conditioned Medium with Different Basal Medium during Mouse Embryo In Vitro Culture.

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

doi:10.3390/ani10081414

Abstract

Simple SummaryAssisted reproductive techniques, which are used to resolve various infertility problems, have advanced following the emphasis on their use. Embryos produced in vitro rather than in vivo are exposed to greater stress, with the quality of the embryos being affected by the in vitro culture conditions. To reduce oxidative stress and consequent apoptosis of embryos for successful implantation and pregnancy maintenance, the present study evaluated the anti-oxidative effect of human adipose stem cell conditioned medium (ASC-CM) with different basal medium as supplement in in vitro culture (IVC) medium for mouse preimplantation embryo. Treatment of 5% human ASC-CM based on Dulbecco′s modified Eagle′s medium (DMEM-CM) indicated an enhanced development of mouse in vitro fertilized embryo, decreased expression level of indicators for oxidative stress, and apoptosis in blastocysts. To our knowledge, this is the first study to demonstrate that DMEM-CM can be an optimal supplement during IVC to promote in vitro embryo development and the success rate of assisted reproduction with its anti-oxidative and anti-apoptotic effects.The quality of embryos produced by assisted reproductive techniques should be advanced by the improvement of in vitro culture conditions for successful implantation and pregnancy maintenance. We investigated the anti-oxidative effect of human adipose stem cell (ASC) conditioned medium with its optimal basal medium, Dulbecco′s modified Eagle′s medium (DMEM-CM), or keratinocyte serum-free medium (KSFM-CM) as supplements during in vitro culture (IVC) of in vitro fertilized mouse embryo. At first, preimplantation embryo development was evaluated in KSFM-CM and DMEM-CM supplemented cultures at various concentrations. The blastocyst (BL) and hatched BL formation rates were significantly increased in 5% DMEM-CM, while no difference was observed from KSFM-CM. Next, comparing the efficacy of KSFM-CM and DMEM-CM at the same concentration, DMEM-CM enhanced the developmental rate of 16 cells, morula, BL, and hatched BL. The expression level of reactive oxygen species decreased and that of glutathione increased in BL cultured with DMEM-CM, which confirms its anti-oxidative effect. Furthermore, apoptosis in BL cultured with DMEM-CM was reduced compared with that in KSFM-CM. This study demonstrated that the comparative effect of human ASC-CM made of two different basal media during mouse embryo IVC and anti-oxidative effect of 5% DMEM-CM was optimal to improve preimplantation embryo development.

Highlights

Infertility, defined as failure to become pregnant despite sexual intercourse without contraception for more than a year, is a prevalent condition among approximately 15% of couples worldwide [1].To resolve the medical, social, and economic concerns that arise from infertility, assisted reproduction has been globalized, resulting in an increase in the birth rate globally [2]
Flow cytometric analysis was conducted to identify the expression of mesenchymal stem cell (MSC)-specific markers in isolated adipose stem cell (ASC) from healthy female donors (Figure 1A–C)
Our results demonstrated that 5% DMEM-conditioned medium (CM) improved preimplantation and keratinocyte serum-free medium (KSFM)-CM groups

Summary

Introduction

Social, and economic concerns that arise from infertility, assisted reproduction has been globalized, resulting in an increase in the birth rate globally [2]. According to the centers for disease control and prevention, the definition of assisted reproductive technologies (ARTs) covers all fertility treatments with the manipulation of gametes and embryos [3], and among ARTs, in vitro culture (IVC) is an essential step for the development of embryos, which can be implanted to result in successful pregnancy. The interaction of growth factors and cytokines from female reproductive tracts with embryos are required for improving the optimal microenvironment during IVC [6], and the supplementation of embryos in IVC medium with various growth factors has shown an effect to reduce apoptosis and degeneration, and increase blastocyst (BL)

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