Stem Cell–Derived Bioactive Materials Accelerate Development of PorcineIn Vitro–Fertilized Embryos

Abstract

Stem cells show the capability to proliferate in an undifferentiated state with long-term self-renewal, which gives the cells advantages for use as bioactive material (BM) for embryo culture in vitro. The objective of this experiment was to investigate the effect of two BMs-human adipose tissue-derived mesenchymal stem cell BM (hAT-MSC-BM) and human embryonic stem cell-derived BM (hESC-BM)-on porcine embryo development compared to commonly used bovine serum albumin (BSA) or serum treatment groups. In vitro-fertilized (IVF) embryos were cultured in PZM-5 with 4 mg/mL BSA until day 4 and equally divided into four groups. Starting from day 4 (until day 6), each group was treated with the following protein additives: 4 mg/mL BSA (control), 10% fetal bovine serum (FBS), 10% hAT-MSC-BM, or 10% hESC-BM. Our results show FBS-and two other BM-treated groups showed significant increases in blastocyst formation rate, hatching rate, and total cell number compared with the control group ( p < 0.05). The hAT-MSC-BM and hESC-BM treatment groups presented better-quality embryo development, especially from the middle expanding stage to hatching. In particular, the hAT-MSC-BM-treated group showed the highest developmental potential of all groups and formed the most expanding-stage blastocysts. The relative expression of reprogramming-related transcription factor (POU5F1, SOX2, DPPA5, and CDH1), antioxidant (PRDX5), and apoptosis (BCL2L1 and BIRC5) genes also increased in two types of BMs compared to the control. In addition, we investigated the protein synthesis of the tight junction-and gap junction-related genes, connexin 43 and zonula occludens-1 (ZO-1); these increased more than in the control. These results demonstrate that stem cell-derived BMs accelerate porcine preimplantation embryo development and that the BMs would be helpful in the development of preimplantation embryos.

Allicin protects porcine oocytes against damage during aging in vitro

“…Western blots were performed essentially as described previously (Lee, Moon, Kim, & Park, ). In brief, oocytes (40 per sample) were solubilized in 20 μl of 1 × sodium dodecyl sulfate (SDS) sample buffer (62.5 mM Tris‐HCl, pH 6.8, containing 2% (w/v) SDS, 10% (v/v) glycerol, 50 μM dithiothreitol, and 0.01% (w/v) bromophenol blue or phenol red) and heated for 5 min at 95°C.…”

“…Western blots were performed essentially as described previously (Lee, Moon, Kim, & Park, 2015). In brief, oocytes ( p values < .05 were considered significant.…”

The antioxidant dieckol reduces damage of oxidative stress‐exposed porcine oocytes and enhances subsequent parthenotes embryo development

“…Treatment with stem cell factor, a growth factor that reduces cellular apoptosis, increases BCL2L1 expression, while antiapoptotic genes are poorly expressed in degenerative embryos (Felici et al, 1999). A recent study reported that treatment of in vitro fertilized porcine oocytes with mesenchymal stem cell‐derived bioactive material (hAT‐MSC‐BM) and human embryonic stem cell‐derived bioactive material (hESC‐BM) increases expression of transcription factors including POU5F1 and SOX2 , as well as antiapoptotic genes including BCL2L1 and BIRC5 (S. E. Lee et al, 2015). Another study reported that TNFAIP2, BAX, POU5F1 , and BCL2L1 are highly expressed in zinc‐treated in vitro fertilized blastocysts (Y. Jeon et al, 2014).…”

Lysophosphatidic acid accelerates development of porcine embryos by activating formation of the blastocoel

“…For example, the addition of γ‐linolenic acid into the medium of bovine in vitro‐produced embryos promoted the developmental competence of zygotes and increased the cleavage rate until Day 2 (Gaja et al, ). Treatment with stem cell‐derived bioactive materials also accelerated the development of porcine in vitro‐ fertilized embryos (Lee, Moon, Kim, & Park, ). We found that rates of blastocyst development and over‐expanded blastocyst development were higher when embryos were treated with LPA starting on Day 4 post‐activation (Figure , Table ).…”