Department of Developmental Biology, Faculty of Basic Sciences and Advanced Technologies in Biology, University of Science and Culture, ACECR, Tehran, Iran; Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
Eur J Obstet Gynecol Reprod Biol. 2021 Mar;258:139-145. doi: 10.1016/j.ejogrb.2020.12.048. Epub 2020 Dec 29.
To preserve human ovarian tissue structure and improve follicular growth and survival during in-situ culture, various biomaterials are used. In this study we aimed to compare agar as a cultivation substrate with matrigel-coated insert in order to achieve an optimum system for in-situ human follicle culture.
Frozen-thawed human ovarian cortical tissues were cultured on either matrigel-coated inserts or agar-soaked substrates. The proportion of morphologically viable and degenerated follicles at different developmental stages, secreted hormonal levels, and apoptotic and proliferation gene expressions were compared between the cultured groups after 7-days of culture.
The follicular growth was not significantly different between the two cultured groups, although showing higher percentage of growing follicles in agar cultured group. The secreted hormonal levels didn't have any difference between two cultured groups. Although the apoptotic gene expressions didn't show any difference between the cultured groups, the apoptotic index was lower in agar cultured group. In addition, Ki67 gene expression, a proliferative marker, showed a significantly higher expression in agar cultured group.
Based on the results, agar is as suitable as matrigel-coated inserts for the survival and growth of follicles during culture. Therefore, agar can be an inexpensive alternative substrate for culturing frozen-thawed human ovarian cortical strips.
为了在原位培养过程中保存人卵巢组织的结构并提高卵泡的生长和存活率，使用了各种生物材料。本研究旨在比较琼脂作为培养基质与基质胶包被的插入物，以实现原位人卵泡培养的最佳系统。
将冷冻-解冻的人卵巢皮质组织分别培养在基质胶包被的插入物或琼脂浸泡的基质上。培养 7 天后，比较两组培养物中不同发育阶段形态上有活力和退化的卵泡的比例、分泌的激素水平以及凋亡和增殖基因的表达。
两组培养物的卵泡生长没有显著差异，尽管琼脂培养组的生长卵泡比例较高。两组培养物的分泌激素水平没有差异。尽管凋亡基因表达在培养组之间没有差异，但琼脂培养组的凋亡指数较低。此外，增殖标志物 Ki67 基因的表达在琼脂培养组中表现出更高的表达。
根据结果，琼脂与基质胶包被的插入物一样适合于培养过程中卵泡的存活和生长。因此，琼脂可以作为培养冷冻-解冻人卵巢皮质带的廉价替代基质。
