Asian Pacific Journal of Reproduction

256-265

Authors: Nguyen Sa Viet Le, Minh Tam Le, Thanh Ngoc Cao

Objective: To compare the effectiveness of inositol and metformin on the clinical characteristics, and endocrine and metabolic profiles of infertile polycystic ovarian syndrome (PCOS) women from Vietnam. Methods: From June 2018 to August 2022, a randomized trial was undertaken at the Hue Center for Endocrinology and Reproduction on infertile women aged 18 to 40 years with polycystic ovarian syndrome. The clinical, endocrine, and metabolic features of these individuals were assessed before and after 3 months of treatment with 2 g of inositol or 1700 mg of metformin per day. Natural pregnancy rates, adverse effects, and tolerance of inositol were recorded. Results: The study included 171 infertile PCOS women who were eligible to participate and took part in the baseline assessment, of whom 132 women participated in data analysis after 3 months. After metformin treatment, 42.1% of women with oligomenorrhea experienced regular menstruation. Metformin significantly lowered body mass index (BMI), waist circumference and testosterone levels, but had no effect on other clinical characteristics, endocrine profiles, or metabolic profiles. 29.2% Of women reported experiencing side effects. 21% Of them attained pregnancy, which resulted in 17.1% of live births. In the inositol group, the rate of regular cycle increased by 18.2% and the total testosterone concentration significantly decreased. In overweight/obese women with PCOS, inositol significantly decreased weight, BMI, waist and hip circumferences (P<0.05). 100% Of women tolerated inositol and continued treatment. 18.9% Of them became pregnant, leading to 17% of live births. Conclusions: Metformin and inositol can improve weight and waist circumference in overweight/obese infertile women with PCOS. Metformin is associated with a higher rate of regular menstruation, whereas inositol is associated with a lower rate of adverse effects. The spontaneous conception, clinical pregnancy, and live birth rates between two groups are comparable.

273-280

Authors: R. Kumar Pramod, Deepthi Varughese, A. Javed Jameel, Bhisma Narayan Panda, Soma Goswami, Abhijit Mitra

Objective: To assess the effect of cryopreservation on stemness and proliferation potential of sheep spermatogonial stem cells (SSCs) in vitro. Methods: Sheep testicular cells were isolated and putative SSCs were enriched by the laminin-based differential plating method. Putative SSCs were co-cultured with the Sertoli cell feeder prepared by the Datura Stramonium Agglutinin (DSA-lectin)-based method. The cultured putative SSCs were cryopreserved in Dulbecco's Modified Eagle Medium-10% fetal bovine serum mixture (DMEM- 10% FBS) media containing 10% dimethyl sulfoxide (DMSO) alone or 10% DMSO plus 200 mM trehalose. Cryopreserved putative SSCs were evaluated for their proliferation potential using in vitro culture and stemness by immunocytochemistry. Finally, the transfection ability of cryopreserved putative SSCs was analyzed. Results: We isolated 91% viable testicular cells from sheep testes. The majority of the laminin enriched cells expressed the SSC related marker, ITGA6. Co-culture of sheep putative SSCs with Sertoli cell feeder resulted in the generation of stable colonies, and the expression of SSC marker was maintained after several passages. A significantly higher number of viable putative SSCs was recovered from SSCs cryopreserved in media containing 10% DMSO and 200 mM trehalose compared to 10% DMSO alone (P<0.01). Cryopreserved putative SSCs formed colonies and showed SSC marker expression similar to the non-cryopreserved putative SSCs. The appearance of green fluorescent colonies over the Sertoli cell feeder indicated that cryopreserved sheep SSCs were successfully transfected. Conclusions: Cryopreserved putative SSCs can retain their stemness, colony forming ability, and transfection efficiency in vitro. Our research may help in the effective preservation of germplasm and the generation of transgenic ovine species.

288-298

Authors: P. Perumal, Jai Sunder, A. K. De, D. Bhattacharya, A. K. Nahak, R. Vikram, E. B. Chakurkar

Objective: To measure seasonal effects on semen quality profiles, seminal biochemical and oxidative stress attributes in fresh and liquid stored semen in monsoon and dry seasons. Methods: A total of 10 Teressa bucks (3-4 years) were selected from breeding farm, ICAR-Central Island Agricultural Research Institute, Port Blair, Andaman and Nicobar Islands, India. Semen samples (n=25 per season) were collected through artificial vagina method and preserved at refrigerated temperature (5 ℃ ) for 48 h using Tris citrate glucose based extender. We detected semen quality parameters [volume, mass activity, pH, sperm concentration, total motility, viability, total sperm abnormality, and plasma membrane, acrosomal and nuclear integrities], biochemical profiles [aspartate amino transferase (AST), alanine amino transferase (ALT) and total cholesterol], and oxidative stress markers [total antioxidant capacity (TAC) and malondialdehyde (MDA)] during monsoon and dry seasons. Results: Semen quality parameters significantly differed between seasons (P<0.05) and among storage periods (P<0.05). Volume, pH, mass activity, motility, viability, acrosomal, plasma membrane and nuclear integrities, and TAC were significantly higher (P<0.05). Sperm concentration, sperm abnormalities, MDA, AST, ALT and total cholesterol were significantly lower in fresh semen of monsoon than dry season (P<0.05). Motility, viability, acrosomal, plasma membrane and nuclear integrities, and TAC were significantly decreased (P<0.05) while sperm abnormality, AST, ALT, total cholesterol and MDA were significantly increased as liquid semen storage period advanced (P<0.05). Conclusions: Monsoon season has higher beneficial effects on semen quality profiles and liquid stored semen remained usable for upto 48 h. Good quality ejaculates with higher TAC and lower MDA can be cryopreserved and will be used for artificial insemination.