Comparison of Morphology and Development of Follicles Derived from Cultured Mouse Ovarian Tissue in Three Methods: Hanging Drop, under Mineral Oil and Insert

Authors
M. Amoushahi 1
M. Salehnia 1
S. J. Mola 2
1 Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2 Department of Molecular Genetics, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Abstract
Objective: The present study compared the efficiency of three mouse ovarian tissue culture methods - hanging drop, under mineral oil, and on the insert with regards to improving in vitro ovarian follicular development.
Methods: Ovaries from 7-day-old old female NMRI mice sacrificed by cervical dislocation were collected and cultured in α-MEM medium supplemented with 5% fetal bovine serum for 7 days in 3 groups (hanging drop, under mineral oil and on the insert). We evaluated and compared the morphology and surface area of the ovaries and percentage of normal follicles in all groups. After the 7-day culture, the ovaries cultured on the insert showed better growth compared to the other groups. Their preantral follicles were isolated and cultured for 12 days. We assessed the follicular diameter, survival and maturation rate of these oocytes at the end of the last culture period.
Results: The percentages of normal follicles in cultured ovaries were 73.85±2.49% (insert), 51.63±3.93% (hanging drop), and 40.52±5.86% (mineral oil) after the 7-day culture. The percentage of preantral follicles significantly increased from 2.1±0.44 to 24.5±2.4 in the group cultured on insert (P<0.05), however there was no significant difference in the other groups (P>0.05). There were significantly increased surface areas of the ovarian tissues after the 7-day culture in all groups (P<0.05). Ovaries cultured on the insert had a diameter of isolated follicles after 12 days of culture of 410±7.07 µm and the MII rate was 30.26%.
Conclusion: The ovarian growth and morphology were well preserved in tissues cultured on the insert compared to the other culture methods.

Keywords

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Pathobiology Reserach
Volume 18, Issue 3
October 2015
Pages 59-73