Katharina Winkler-Crepaz1, Verena Nederegger1, Sarrah Ayuandari2, Doris Rosenfellner1, Ioannis Zervomanolakis3, Susanne Hofer1, Ludwig Wildt1, Stephanie C Ziehr4. 1. Department of Gynecological Endocrinology and Reproductive Medicine, Innsbruck Medical University, Innsbruck, Austria. 2. Department of Gynecological Endocrinology and Reproductive Medicine, Innsbruck Medical University, Innsbruck, Austria; Department of Obstetrics and Gynecology, Gadjah Mada University, Yogyakarta, Indonesia. 3. Department of Gynecological Endocrinology and Reproductive Medicine, Innsbruck Medical University, Innsbruck, Austria; Mitera IVF, Mitera Assisted Reproduction Unit, Athens, Greece. 4. Department of Gynecological Endocrinology and Reproductive Medicine, Innsbruck Medical University, Innsbruck, Austria. Electronic address: stephanie.ziehr@i-med.ac.at.
Abstract
OBJECTIVE: To evaluate the impact of dynamic in vitro culture on initiation of early follicular growth in prepubertal mouse ovaries. DESIGN: Ovaries from 8-day-old BALB/c mice were cultured either in a dynamic system (n=28) or in a static system (n=20) for 4 days. Uncultured 8-day-old (n=9) or 12-day-old (n=17) ovaries served as baseline or in vivo controls, respectively. SETTING: Academic research center. ANIMAL(S): Newborn female BALB/c mice (n=37). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Histologic follicle classification and counting and assessment of follicular viability via immunofluorescent staining. RESULT(S): The percentage of secondary follicles after dynamic culture was identical to the 12-day-old in vivo control. In contrast, after static culture ovaries showed a significantly higher percentage of secondary follicles. For immunofluorescent viability assessment 6.78 follicles per ovary could be isolated after dynamic culture, whereas only 3.8 follicles per ovary could be isolated after static culture. CONCLUSION(S): Dynamic in vitro culture supports physiologic follicular growth initiation, comparable to that observed in vivo. In contrast, accelerated follicular growth was observed after static culture. These findings add additional evidence to the idea that dynamic culture might be a beneficial first step to initiate follicle growth in vitro within the context of fertility preservation.
OBJECTIVE: To evaluate the impact of dynamic in vitro culture on initiation of early follicular growth in prepubertal mouse ovaries. DESIGN: Ovaries from 8-day-old BALB/c mice were cultured either in a dynamic system (n=28) or in a static system (n=20) for 4 days. Uncultured 8-day-old (n=9) or 12-day-old (n=17) ovaries served as baseline or in vivo controls, respectively. SETTING: Academic research center. ANIMAL(S): Newborn female BALB/c mice (n=37). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Histologic follicle classification and counting and assessment of follicular viability via immunofluorescent staining. RESULT(S): The percentage of secondary follicles after dynamic culture was identical to the 12-day-old in vivo control. In contrast, after static culture ovaries showed a significantly higher percentage of secondary follicles. For immunofluorescent viability assessment 6.78 follicles per ovary could be isolated after dynamic culture, whereas only 3.8 follicles per ovary could be isolated after static culture. CONCLUSION(S): Dynamic in vitro culture supports physiologic follicular growth initiation, comparable to that observed in vivo. In contrast, accelerated follicular growth was observed after static culture. These findings add additional evidence to the idea that dynamic culture might be a beneficial first step to initiate follicle growth in vitro within the context of fertility preservation.