Scholarlink

<p>In the present study, germline stem cells were isolated from 6-month-old gonads of <i>Huso huso</i> using different concentrations of trypsin and collagenase. The resulting cells were cultured in Leibovitz’s L-15 (L-15) and Dulbecco’s Modified Eagle Medium (DMEM) for 3&#xa0;weeks. The identity of germline stem cells was confirmed by immunocytochemical staining using a VASA-specific antibody. The expression of <i>Vasa</i>, <i>Nanos</i>, <i>Bax,</i> and <i>p53</i>-<i>like</i> genes was assessed using qRT-PCR. Cell cryopreservation was also performed using dimethyl sulfoxide (DMSO) and ethylene glycol (EG). Based on results, enzymatic digestion with 0.5% trypsin–EDTA combined with 1&#xa0;mg/ml collagenase was the most effective isolation method. In comparison of culture media, L-15 supported the formation of larger colonies (101.87 ± 8.40&#xa0;μm), while DMEM produced a greater number of colonies. <i>Vasa</i> and <i>Nanos</i> gene expression reached its highest level on the 20th day of culture (1.54 and 2.23-fold, respectively), while <i>Bax</i> and <i>p53-like</i> expression significantly increased on the 33rd day. Also, DMSO showed better performance in freezing cells compared to EG and maintained 67% cell recovery after thawing. Overall, optimizing the isolation, culture, and cryopreservation of germline stem cells could be used as a valuable tool in conservation and recovery programs for the endangered <i>Huso huso.</i></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Characterization and preservation of germline stem cells in endangered beluga sturgeon (Huso huso) gonads: implications for conservation

  • 期刊论文

  • 发表日期：

    2026年7月19日

查看全文
  • Hediye Fadakar,
  • Tooba Mirzapour,
  • Shirin Jamshidi,
  • Tooraj Sohrabi

摘要

In the present study, germline stem cells were isolated from 6-month-old gonads of Huso huso using different concentrations of trypsin and collagenase. The resulting cells were cultured in Leibovitz’s L-15 (L-15) and Dulbecco’s Modified Eagle Medium (DMEM) for 3 weeks. The identity of germline stem cells was confirmed by immunocytochemical staining using a VASA-specific antibody. The expression of Vasa, Nanos, Bax, and p53-like genes was assessed using qRT-PCR. Cell cryopreservation was also performed using dimethyl sulfoxide (DMSO) and ethylene glycol (EG). Based on results, enzymatic digestion with 0.5% trypsin–EDTA combined with 1 mg/ml collagenase was the most effective isolation method. In comparison of culture media, L-15 supported the formation of larger colonies (101.87 ± 8.40 μm), while DMEM produced a greater number of colonies. Vasa and Nanos gene expression reached its highest level on the 20th day of culture (1.54 and 2.23-fold, respectively), while Bax and p53-like expression significantly increased on the 33rd day. Also, DMSO showed better performance in freezing cells compared to EG and maintained 67% cell recovery after thawing. Overall, optimizing the isolation, culture, and cryopreservation of germline stem cells could be used as a valuable tool in conservation and recovery programs for the endangered Huso huso.