文献类型：会议论文

英文题名：In vitro production of haploid germ cells from murine spermatogonial stem cells using a two-dimensional cell culture system

作者：Ibtisham F.[1,2];Zhao Y.[1];Nawab A.[1];Wu J.[1];Mei X.[1];Honaramooz A.[2];An L.[1]

机构：[1]College of Agriculture, Guangdong Ocean University, Zhanjiang, Guangdong, China;[2]Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, SK, Canada

语种：英文

外文关键词：Haploid male germ cells; In vitro propagation; In vitro spermatogenesis; Infertility treatment; Retinol; Spermatogonial stem cells (SSCs)

外文摘要：The in vitro propagation and differentiation of spermatogonial stem cells (SSCs) has many potential applications within reproductive science and medicine. We established a two-dimensional (2D) cell culture system to proliferate and differentiate prepubertal mouse SSCs as a model capable of maximizing on a small number of donor SSCs. We also investigated the effects of retinol on in vitro SSC differentiation. Testis cells were cultured for 10 days in a serum-free medium. This produced SSC colonies which were then dissociated and sub-cultured for an additional 20 days in a differentiation medium. Before inducing differentiation, colonies expressed genes specific for undifferentiated spermatogonia (Ngn3, Plzf). After 10 days in the differentiation medium, Stra8 expression was upregulated. After 20 days, Acr expression was upregulated, indicating the completion of meiosis. Immunofluorescence, RT-PCR and flow cytometry confirmed the presence of haploid male germ cells (4.4% of all cells). When retinol was added to the differentiation medium the proportion of haploid germ cells increased (8.1% of cells). We concluded that, under serum-free culture conditions, prepubertal SSCs will generate colonies that can differentiate into haploid germ cells in a 2D culture system. These cells demonstrate a relatively high efficiency of haploid-cell production, which can be further improved with retinol. ? 2021 Elsevier Inc.