Development of an in vitro culture system for immature oocytes of red swamp crayfish (Procambarus clarkii)
文献类型： 外文期刊
作者： Shui, Yan 1 ; Zhan, Ming 2 ; Li, Jiajia 3 ; Su, Shengyan 1 ;
作者机构： 1.Chinese Acad Fishery Sci, Freshwater Fisheries Res Ctr, Key Lab Integrated Rice Fish Farming Ecol, Minist Agr & Rural Affairs, Wuxi 214081, Peoples R China
2.Nanjing Agr Univ, Wuxi Fisheries Coll, Wuxi 214081, Peoples R China
3.Jiangsu Inst Freshwater Fisheries, Nanjing 210017, Peoples R China
关键词： Procambarus clarkii; Ovarian tissue; Oocytes; In vitro culture; 17-OHP
期刊名称：ANIMAL REPRODUCTION SCIENCE （ 影响因子：2.22； 五年影响因子：2.315 ）
ISSN： 0378-4320
年卷期： 2022 年 243 卷
页码：
收录情况： SCI
摘要： The present study assessed general strategies for the isolation and in vitro culture of immature ovary-derived cells (oocytes) in red swamp crayfish (Procambarus clarkii). In phase 1 of this study, the effects of different digestive enzymes on cell retrieval from developing ovarian tissues were examined to facilitate efficient oocyte isolation. Subsequently, the ovary-derived cells/oocytes were cultured under various conditions to evaluate the effects of basal media and supplement concentrations. Collagenase I treatment at 26 celcius for 25 min was more effective (approx. 95 %) in oocytes isolation from ovarian tissues than other treatments. Furthermore, DMEM media with HEPES (DMEM-H) were more effective than L15 for oocyte culture (72.4 % survival rate), while oocyte survival improved to 90.6 % using a 10 % fetal bovine serum supplement. In phase 2, the top-performing media were used in immature oocyte culture to assess 17 alpha-Hydroxyprogesterone (17-OHP) induction in vitro. An up-regulated expression of maturity-related genes, i.e., cyclin B, cdc2, ef1 alpha, after 12 h of incubation was evident in qPCR results, demonstrating that the exogenous hormone 17-OHP could induce P. clarkii oocyte maturation in vitro. These results may provide a basis for developing an in vitro system for P. clarkii germline cell culture, which may ultimately lead to the genetic improvement of this species.
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