Location: Warmwater Aquaculture Research Unit
Title: Effect of cryopreservation techniques on proliferation and apoptosis of cultured equine ovarian tissueAuthor
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GASTAL, GUSTAVO - Southern Illinois University |
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AGUIAR, FRANCISCO - Southern Illinois University |
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ISHAK, GHASSAN - Southern Illinois University |
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CAVINDER, CLAY - Mississippi State University |
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WILLARD, SCOTT - Mississippi State University |
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RYAN, PETER - Mississippi State University |
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FEUGANG, JEAN - Mississippi State University |
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GASTAL, EDUARDO - Southern Illinois University |
Submitted to: Theriogenology
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 11/30/2018 Publication Date: 12/1/2018 Citation: Gastal, G., Aguiar, F., Ishak, G., Cavinder, C., Willard, S., Ryan, P., Feugang, J., Gastal, E. 2018. Effect of cryopreservation techniques on proliferation and apoptosis of cultured equine ovarian tissue. Theriogenology. 126:88-94. https://doi.org/10.1016/j.theriogenology.2018.11.034. DOI: https://doi.org/10.1016/j.theriogenology.2018.11.034 Interpretive Summary: Preservation of cellular integrity and its mechanisms after ovarian tissue cryopreservation (OTC) and in vitro culture (IVC) procedures are crucial aspects for the success of preservation and recovery of female fertility. This study aimed to evaluate the effects of two cryopreservation methods (slow-freezing, SF, and vitrification, VIT) on the equine ovarian tissue after 1, 3, and 7 days of IVC by assessing: (i) preantral follicle morphology and distribution of follicle classes; (ii) protein expression of markers of cell proliferation for EGFR and Ki-67; (iii) markers of apoptosis for Bax and Bcl-2; and (iv) DNA fragmentation. Obtained results shed light on equine fertility preservation programs using OTC techniques. Technical Abstract: The percentages of normal primordial follicles were similar (P > 0.05) among SF-control, VIT-control, and fresh control groups. After 7 days of culture, VIT-IVC7 had a greater (P < 0.05) total percentage of normal preantral follicles when compared with SF-IVC7, but both had a lower (P < 0.05) percentage than fresh IVC7 group. Prior to and after 7 days of culture, expression of EGFR and Ki-67 were similar (P > 0.05) among fresh, SF, and VIT groups. After 7 days of culture, VIT had higher (P < 0.05) Bax expression than the fresh and SF tissues, but Bcl-2 was similar (P > 0.05) among groups. Prior to IVC, TUNEL signals were similar (P > 0.05) among groups; however, VIT-IVC7 had greater (P < 0.05) TUNEL signals when compared with the fresh IVC7 group. In conclusion, findings demonstrated: (i) similar efficiency between SF and VIT compared with fresh control to preserve morphologically normal follicles; and (ii) similar tissue functionality and cell proliferation capability after equine OTC by either SF and VIT methods following IVC for 7 days. |