Effect of various co-culture systems on embryo development in ovine

Abstract

Considering the advent of mesenchymal stem cells (MSCs) as a new source of somatic cells in embryo co-culture system, the current study was aimed to compare in vitro embryo development using embryonic MSCs monolayer with embryonic fibroblast cells (EFCs), oviductal epithelial cells (OECs), and cell-free culture system. The IVM/IVF presumptive sheep zygotes were randomly cultured in different culture conditions as follows: (1) SOFaaBSA medium for the whole culture period (SOF, n = 371), (2) SOFaaBSA medium for the first 3 days followed by co-culturing with MSCs for the next 5 days (SOF-MSCs, n = 120), (3) co-culturing with MSCs for the first 3 days followed by culture in SOFaaBSA medium for the next 5 days (MSCs-SOF, n = 133), (4) co-culturing with MSCs for the whole culture period (MSCs, n = 212), (5) SOFaaBSA medium for the first 3 days followed by co-culturing with EFCs for the next 5 days (SOF-EFCs, n = 132), (6) co-culturing with EFCs for the first 3 days followed by culture in SOFaaBSA medium for the next 5 days (EFCs-SOF, n = 165), (7) co-culturing with EFCs for the whole culture period (EFCs, n = 236), and (8) co-culturing with OECs for the whole culture period (OECs, n = 255). One-Way ANOVA by multiple pairwise comparisons using Tukey's test was performed. Co-culturing in MSCs group had no superiority over EFCs and OECs groups. Though, when co-culturing with MSCs and EFCs was limited to the first 3 days of culture, the embryo development indices were improved compared to the other co-cultured groups. Considering both the hatching rate and total cell number, the application of MSCs for the first 3 days of culture (MSCs-SOF) was superior to the other co-culture and SOF groups.

Human amniotic membrane stem cells’ conditioned medium has better support for in‐vitro production of bovine embryos than FBS

“…The trophic effect of CM may be due to the presence of antioxidants and growth factors in CM compared to the control group as such the number of ICM and total cells in 5% CM group was approximately 2 and 1.6 times of control group, respectively (Table 3). These results were consistent with the results of some other researchers (Bhardwaj et al., 2016; Heidari et al., 2013). In buffalo, the presence of LIF in stem cell‐derived condition media had a significant effect on blastocyst development, hatching rate and total number of cells in blastocyst (Bhardwaj et al., 2016).…”

Chemobrionic structures in tissue engineering: self-assembling calcium phosphate tubes as cellular scaffolds

“…Stem cells are multipotent progenitors that are capable of differentiating down several specialised cell lineages, including that of bone, cartilage and cardiac cell types, making them highly valuable for applications in tissue engineering. 40,41 Given that cellular adhesion is crucial for cell differentiation, migration and survival, demonstrating the adherence of BMSCs to chemobrionic scaffolds is a pivotal first step toward validating the future developments of materials. 42 In order to assess cell viability, scaffold BMSC cultures were stained with NucBlue® Live and NucGreen® Dead reagents and imaged using fluorescence microscopy.…”

Novelties in Ovine Assisted Reproductive Technologies – A Review

“…Following 0.5-1-hour incubation at 38.5 °C, the upper level of the medium (0.5-1 ml) is collected and centrifuged (200 x g) for 10 minutes. The Percoll gradient method is used for frozenthawed ram sperm (73,74). The Percoll granules are composed of colloidal silica (15-30 nm) coated with polyvinylpyrrolidone.…”