Increasing of blastocyst rate and gene expression in co-culture of bovine embryos with adult adipose tissue-derived mesenchymal stem cells

Abstract

Purpose Despite advances in the composition of defined embryo culture media, co-culture with somatic cells is still used for bovine in vitro embryo production (IVEP) in many laboratories worldwide. Granulosa cells are most often used for this purpose, although recent work suggests that co-culture with stem cells of adult or embryonic origin or their derived biomaterials may improve mouse, cattle, and pig embryo development. Materials and methods In experiment 1, in vitro produced bovine embryos were co-cultured in the presence of two concentrations of bovine adipose tissue-derived mesenchymal cells (b-ATMSCs; 10 3 and 10 4 cells/mL), in b-ATMSC preconditioned medium (SOF-Cond), or SOF alone (control). In experiment 2, co-culture with 10 4 b-ATMSCs/mL was compared to the traditional granulosa cell co-culture system (Gran). Results In experiment 1, co-culture with 10 4 b-ATMSCs/mL improved blastocyst rates in comparison to conditioned and control media (p < 0.05). Despite that it did not show difference with 10 3 b-ATMSCs/mL (p = 0.051), group 10 4 bATMSCs/mL yielded higher results of blastocyst production.In experiment 2, when compared to group Gran, co-culture with 10 4 b-ATMSCs/mL improved not only blastocyst rates but also quality as assessed by increased total cell numbers and mRNA expression levels for POU5F1 and G6PDH (p < 0.05). Conclusions Co-culture of bovine embryos with b-ATMSCs was more beneficial than the traditional co-culture system with granulosa cells. We speculate that the microenvironmental modulatory potential of MSCs, by means of soluble substances and exosome secretions, could be responsible for the positive effects observed. Further experiments must be done to evaluate if this beneficial effect in vitro also translates to an increase in offspring following embryo transfer. Moreover, this study provides an interesting platform to study the basic requirements during preimplantation embryo development, which, in turn, may aid the improvement of embryo culture protocols in bovine and other species.

Protein Cargo of Extracellular Vesicles From Bovine Follicular Fluid and Analysis of Their Origin From Different Ovarian Cells

“…In reproductive fluids, Almiñana et al showed that mRNAs encoding ribosomal proteins of the large (RPL) and small subunit (RPS) were the most abundant protein-coding RNAs within oviductal EVs ( 34 ). These results are in line with studies in EVs from other body fluids including saliva ( 58 ) and urine ( 59 ) It is possible that the numerous ribosomal proteins identified in our ffEV preparations, and those identified by others, may form RNA–ribonucleoprotein complexes that function to package RNAs into EVs and, thus, participate in RNA maintenance ( 57 ) or RNA degradation in exosomes. The presence of numerous RBPs and ribosomal proteins in ffEVs may suggest a possible role for the proteins in RNA sorting, which was also reported in EVs of different origins ( 60 ).…”

Applications of mesenchymal stem cell technology in bovine species

“…Of the cell passages examined in this study, passage 5 seemed to be the most efficient in the performance of nuclear transfer due to its high level of stemness, multipotency, and the low level of chromatin compaction [76]. The embryo production rate was also shown to improve when embryos were co-cultured with MSC [77], representing in yet another way the importance of MSC in addressing commercial goals.…”

Mesenchymal stem cell: Basic research and potential applications in cattle and buffalo

“…The higher cell concentration (10 4 cells/ml) may be more favorable than lower (10 3 cells/ml) cell concentration and conditioned medium, though blastocyst rates may be more (Miranda et al, 2016).…”

“…In vitro fertilized embryos on culturing with bAD‐MSCs (10 3 /10 4 cells/ml) may show better blastocyst rates and quality as determined by POU5F1 and G6PDH. The higher cell concentration (10 4 cells/ml) may be more favorable than lower (10 3 cells/ml) cell concentration and conditioned medium, though blastocyst rates may be more (Miranda et al, ).…”