文献类型：期刊文献

英文题名：Optimal isolation, culture, and in vitro propagation of spermatogonial stem cells in Huaixiang chicken

作者：Ibtisham, Fahar[1,2];Tang, Shuyan[1];Song, Yiping[1];Wanze, Wang[2];Xiao, Mei[1];Honaramooz, Ali[2];An, Lilong[1]

机构：[1]Guangdong Ocean Univ, Agr Coll, Zhanjiang 524088, Guangdong, Peoples R China;[2]Univ Saskatchewan, Western Coll Vet Med, Dept Vet Biomed Sci, Saskatoon, SK, Canada

年份：2024

卷号：59

期号：7

外文期刊名：REPRODUCTION IN DOMESTIC ANIMALS

收录：SCI-EXPANDED(收录号：WOS:001268313700001)、、Scopus(收录号：2-s2.0-85197779875)、WOS

基金：This work was supported by the Science and Technology Planning Project of Guangdong Province, China (2010 B090400376) and the Natural Sciences and Engineering Research Council of Canada (# 404908).

语种：英文

外文关键词：growth factors; Huaixiang chicken; spermatogonial stem cells; SSC isolation; in vitro culture; SSC purification

外文摘要：Spermatogonial stem cells (SSCs) comprise the foundation of spermatogenesis and hence have great potential for fertility preservation of rare or endangered species and the development of transgenic animals and birds. Yet, developing optimal conditions for the isolation, culture, and maintenance of SSCs in vitro remains challenging, especially for chicken. The objectives of this study were to (1) find the optimal age for SSC isolation in Huaixiang chicken, (2) develop efficient protocols for the isolation, (3) enrichment, and (4) culture of isolated SSCs. In the present study, we first compared the efficiency of SSC isolation using 11 different age groups (8-79 days of age) of Huaixiang chicken. We found that the testes of 21-day-old chicken yielded the highest cell viability. Next, we compared two different enzymatic combinations for isolating SSCs and found that 0.125% trypsin and 0.02 g/L EDTA supported the highest number and viability of SSCs. This was followed by investigating optimal conditions for the enrichment of SSCs, where we observed that differential plating had the highest enrichment efficiency compared to the Percoll gradient and magnetic-activated cell sorting methods. Lastly, to find the optimal culture conditions of SSCs, we compared adding different concentrations of foetal bovine serum (FBS; 2%, 5%, 7%, and 10%) and different concentrations of GDNF, bFGF, or LIF (5, 10, 20, or 30 ng/mL). We found that a combination of 2% FBS and individual growth factors, including GDNF (20 ng/mL), bFGF (30 ng/mL), or LIF (5 ng/mL), best supported the proliferation and colony formation of SSCs. In conclusion, SSCs can be optimally isolated through enzymatic digestion from testes of 21-day-old chicken, followed by enrichment using differential plating. Furthermore, adding 2% FBS and optimized concentrations of GFNF, bFGF, or LIF in the culture promotes the proliferation of chicken SSCs.