Varied Approach of Using MSCs for Bovine Embryo In Vitro Culture

ABSTRACT

The objective of the present study was to evaluate the effect of porcine mesenchymal stem cells (MSCs) secreted factors on bovine in vitro embryo development using MSCs in different culture systems: SOF medium, SOF medium conditioned by MSCs in monolayer, and in reverse drop and by embryo culture in coculture with MSCs. Statistically highly significant differences were noted between the number of blastocysts derived cultures in all tested culture systems. The in vitro culture in SOF turned out to be the most optimal. Statistically highly significant differences were observed in the number of blastocyst obtained between SOF and SOF in coculture with MSCs (P < 0.0001), and between SOF and SOF conditioned (monolayer and drop) (P < 0.00001). The trials to produce blastocysts in SOF conditioned by MSCs in reverse drops and monolayer failed. The blastocysts were obtained and analyzed by TUNEL only in two out of four experimental groups: SOF and SOF in coculture with MSCs. There were no significant differences between any of analyzed blastocysts’ groups neither in the total number of nuclei nor in the apoptotic features. Neither medium conditioning by MSCs in monolayer and in reverse drop nor embryo culture in coculture with MSC turned out to be effective.

KEYWORDS:

• Bovine
• embryo in vitro culture
• medium conditioning
• MSC

Acknowledgments

The authors would like to thank Jarosław Wieczorek Vet Dr for bone marrow aspiration under general anesthesia, and Lechoslaw Gajda for assistance in statistical analysis.

Conflict of interest

None of the authors have any conflict of interest to declare.

Author contributions

This article has not been submitted anywhere else for publication. JO, conception, design of study, interpretation of data; JO and BB, article preparation; JO, BB, and JR, IVM, IVF, IVC; BB and JR, TUNEL; JO and JR, isolation and in vitro culture of MSC as well as medium conditioning by MSC. JR, statistical analysis. All co-authors approved the final draft of the article.

Additional information

Funding

This work was supported by the National Science Centre allocated based on the decision-DEC number 2014/15/B/NZ9/04288, and by National Research Institute of Animal Production statutory activity no. 02-013.1