Wake Forest Institute for Regenerative Medicine (WFIRM), Wake Forest School of Medicine, Winston-Salem, NC 27101, USA.
Facultad de Medicina Universidad de Barcelona, 08036 Barcelona, Spain.
Int J Mol Sci. 2021 Dec 24;23(1):173. doi: 10.3390/ijms23010173.
Klinefelter syndrome (KS) is characterized by a masculine phenotype, supernumerary sex chromosomes (usually XXY), and spermatogonial stem cell (SSC) loss in their early life. Affecting 1 out of every 650 males born, KS is the most common genetic cause of male infertility, and new fertility preservation strategies are critically important for these patients. In this study, testes from 41, XXY prepubertal (3-day-old) mice were frozen-thawed. Isolated testicular cells were cultured and characterized by qPCR, digital PCR, and flow cytometry analyses. We demonstrated that SSCs survived and were able to be propagated with testicular somatic cells in culture for up to 120 days. DNA fluorescent in situ hybridization (FISH) showed the presence of XXY spermatogonia at the beginning of the culture and a variety of propagated XY, XX, and XXY spermatogonia at the end of the culture. These data provide the first evidence that an extra sex chromosome was lost during innate SSC culture, a crucial finding in treating KS patients for preserving and propagating SSCs for future sperm production, either in vitro or in vivo. This in vitro propagation system can be translated to clinical fertility preservation for KS patients.
克氏综合征（KS）的特征是男性表型、额外的性染色体（通常为 XXY）和生精干细胞（SSC）在其早期丧失。KS 影响每 650 名出生男性中的 1 名，是男性不育最常见的遗传原因，对于这些患者，新的生育保存策略至关重要。在这项研究中，对 41, XXY 青春期前（3 天大）的小鼠的睾丸进行了冷冻-解冻。分离的睾丸细胞通过 qPCR、数字 PCR 和流式细胞术分析进行培养和特征分析。我们证明 SSCs 可以在培养物中与睾丸体细胞一起存活并增殖，最多可达 120 天。DNA 荧光原位杂交（FISH）显示，在培养开始时存在 XXY 精原细胞，在培养结束时存在多种增殖的 XY、XX 和 XXY 精原细胞。这些数据首次提供了额外的性染色体在固有 SSC 培养过程中丢失的证据，这是治疗 KS 患者以保存和增殖 SSC 以用于未来的精子产生的关键发现，无论是体外还是体内。这种体外增殖系统可转化为 KS 患者的临床生育保存。
