title: In vitro culture of equine preantral follicles obtained via the Biopsy Pick-Up method.
abstract: The objective was to conduct a preliminary evaluation of the efficacy of two media for in vitro culture of equine preantral follicles. Ovarian cortical strips were obtained from mares (N = 10) via the Biopsy Pick-Up method during the breeding season. Ovarian tissue was immediately submitted to histological analysis (noncultured control; D0) or cultured in situ for 1 day (D1) or 7 days (D7) in either α-MEM or TCM-199 and submitted to histological analysis, generating five treatment groups: noncultured control, α-MEM:D1, TCM-199:D1, α-MEM:D7, and TCM-199:D7. Preantral follicles were evaluated for follicle class (primordial, transitional, primary, and secondary) and morphology (normal vs. abnormal). A total of 142 preantral follicles were analyzed in five replicates. No follicles were observed in the TCM-199:D7 treatment group. The proportion of primordial follicles was higher (P < 0.03) in the control compared to the α-MEM:D7 treatment group. The proportion of primary follicles was higher (P < 0.04) in the α-MEM:D7 treatment group compared to the control. The proportion of developing follicles (transitional, primary, and secondary) was higher (P < 0.03) in the α-MEM:D7 treatment group compared to the control group. There was a greater (P < 0.004) percentage of morphologically normal developing follicles in the α-MEM:D1 treatment group compared to the TCM-199:D1 treatment group. Overall, the percentage of morphologically normal follicles was higher in the control group (72%; P < 0.02) and α-MEM:D1 group (84%; P < 0.0001) compared to the α-MEM:D7 (27%) treatment group. Mean follicle diameter was greater (P < 0.04) in the α-MEM:D7 treatment group (40.6 ± 1.1 μm) compared to the control group (37.3 ± 0.7 μm). Mean oocyte diameter was greater in the α-MEM:D1 (31.0 ± 0.7 μm; P < 0.006), TCM-199:D1 (30.7 ± 1.8 μm; P < 0.006), and α-MEM:D7 (33.2 ± 1.8 μm; P < 0.006) treatment groups compared to the control group (27.4 ± 0.9 μm). In conclusion, based on these preliminary data, in vitro culture of equine ovarian fragments obtained in vivo via the Biopsy Pick-Up method promoted preantral follicle development and follicle and oocyte growth in α-MEM for 7 days, with some follicles remaining morphologically normal throughout the culture period.
authors: Department of Animal Science, Food and Nutrition, Southern Illinois University, Carbondale, Illinois, USA.
authors: K T
authors: KT
authors: Haag
authors: D M
authors: DM
authors: Magalhães-Padilha
authors: G R
authors: GR
authors: Fonseca
authors: A
authors: A
authors: Wischral
authors: M O
authors: MO
authors: Gastal
authors: S S
authors: SS
authors: King
authors: K L
authors: KL
authors: Jones
authors: J R
authors: JR
authors: Figueiredo
authors: E L
authors: EL
authors: Gastal
mesh_terms: D000818:Animals; D001706:Biopsy / Q000379:methods / Q000662:veterinary; D018929:Cell Culture Techniques / Q000662:veterinary*; D003470:Culture Media; D005260:Female; D006736:Horses / Q000502:physiology*; D006080:Ovarian Follicle / Q000166:cytology* / Q000254:growth & development
chemical_list: D003470:Culture Media