Human Adipose-Derived Stem Cells' Paracrine Factors in Conditioned Medium Can Enhance Porcine Oocyte Maturation and Subsequent Embryo Development.

Seok Hee Lee

doi:10.3390/ijms22020579

Abstract

An essential requirement for the success of in vitro maturation (IVM) of the oocyte is to provide an optimal microenvironment similar to in vivo conditions. Recently, somatic cell-based coculture or supplementation of a conditioned medium during IVM has been performed to obtain better quality of oocytes, because they mimic the in vivo reproductive tract by secreting paracrine factors. In this study, human adipose-derived stem cells (ASC) and their conditioned medium (ASC-CM) were applied to IVM of porcine oocytes to evaluate the effectiveness of ASC on oocyte development and subsequent embryo development. In results, both ASC and ASC-CM positively influence on oocyte maturation and embryo development by regulating growth factor receptors (VEGF, FGFR, and IGFR), apoptosis (BCL2), cumulus expansion (PTGS2, HAS2, and TNFAIP6), and oocyte maturation-related genes (GDF9 and BMP15). In particular, the fluorescence intensity of GDF9 and BMP15 was markedly upregulated in the oocytes from the ASC-CM group. Furthermore, significantly high levels of growth factors/cytokine including VEGF, bFGF, IGF-1, IL-10, and EGF were observed in ASC-CM. Additionally, the ASC-CM showed active scavenging activity by reducing the ROS production in a culture medium. Consequently, for the first time, this study demonstrated the effect of human ASC-CM on porcine oocyte development and the alteration of mRNA transcript levels in cumulus–oocyte complexes.

Highlights

Oocyte in vitro maturation (IVM) is a promising assisted reproduction technique (ARTs) for studying embryo development, animal model research, and the treatment of infertility [1,2,3]
The proportion of COCs of COCs exhibiting complete cumulus expansion was significantly increased in adipose-derived stem cells (ASC) coexhibiting complete cumulus expansion was significantly increased in ASC coculture and culture and ASC-conditioned medium (CM) groups compared with the control (Figure 2a–d)
The present study suggests that the coculture ASC with porcine oocytes or the supThe present study suggests that the coculture withmarkedly porcine oocytes or the plementation of Adipose-Derived Stem Cells Conditioned Medium (ASC-CM)

Summary

Introduction

Oocyte in vitro maturation (IVM) is a promising assisted reproduction technique (ARTs) for studying embryo development, animal model research, and the treatment of infertility [1,2,3]. An essential requirement for the success of IVM of the mammalian oocyte is to provide a rich microenvironment for oocyte culture, mimicking the in vivo reproductive tract [4,5]. Somatic cell-based coculture systems [4,11] to obtain a better quality of oocytes. The somatic cell coculture system removes reactive oxygen species (ROS) and provides paracrine factors, which gives beneficial effects on oocyte development [12]. Previous studies have shown that the application of a cell-based coculture system can mimic the in vivo microenvironment by secreting various kinds of growth factors [13,14,15]. Since there are numerous kinds of factors such as cytokines and growth factors, applying all these factors into an IVM system is rather high-priced. The coculture cells still have the possibility to be contaminated until applying to IVM

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