Characterization of rodent Sertoli cell primary cultures.
Characterization of rodent Sertoli cell primary cultures.
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DOI：
10.1002/mrd.23402
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发表时间：
2020-08
影响因子：
2.5
通讯作者：
Reddi PP
中科院分区：
文献类型：
--
作者：
Zomer HD;Reddi PP
支持细胞为雄性生殖细胞的分化提供生理和营养支持，在精子发生过程中起着至关重要的作用。它们形成血睾丸屏障并分泌生殖细胞分化所必需的生长因子。支持细胞原代培养对于理解精子发生的调控至关重要;然而，获得纯培养物一直是一个挑战。啮齿动物支持细胞分离方案不排除间质或结缔组织细胞的污染。支持细胞特异性标记物可能有所帮助，但尚未达成共识。Vimentin是最常用的标记物，并不是支持细胞的特异性标记物，因为已有报道称其在小管周围肌样细胞、间充质干细胞、成纤维细胞、巨噬细胞和内皮细胞中表达，这些细胞会污染支持细胞制剂。基于转录因子和生长因子的标记也有局限性。因此，获得纯支持细胞培养的障碍与分离方法和标记物的使用有关。本综述的目的是讨论当前啮齿动物支持细胞原代培养方法的改进，评估青春期前和成熟支持细胞培养的特性，并提出改进细胞表征的步骤。使用现代方法的潜在好处，包括谱系追踪，特异性细胞消融和RNA-seq获得sertoli特异性转录标记进行了讨论。评估这些标记在蛋白质水平上的特异性和适用性，以表征培养中的支持细胞将是至关重要的。这一综述有望对今后使用啮齿动物支持细胞原代培养的工作产生积极影响。
Sertoli cells play a vital role in spermatogenesis by offering physical and nutritional support to the differentiating male germ cells. They form the blood-testis barrier and secrete growth factors essential for germ cell differentiation. Sertoli cell primary cultures are critical for understanding the regulation of spermatogenesis; however, obtaining pure cultures has been a challenge. Rodent Sertoli cell isolation protocols do not rule out contamination by the interstitial or connective tissue cells. Sertoli cell-specific markers could be helpful, but there is no consensus. Vimentin, the most commonly used marker, is not specific for Sertoli cells since its expression has been reported in peritubular myoid cells, mesenchymal stem cells, fibroblasts, macrophages, and endothelial cells, which contaminate Sertoli cell preparations. Markers based on transcription and growth factors also have limitations. Thus, the impediment to obtaining pure Sertoli cell cultures pertains to both the method of isolation and marker usage. The aim of this review is to discuss improvements to current methods of rodent Sertoli cell primary cultures, assess the properties of prepubertal versus mature Sertoli cell cultures, and propose steps to improve cellular characterization. Potential benefits of using contemporary approaches, including lineage tracing, specific cell ablation, and RNA-seq for obtaining Sertoli-specific transcript markers are discussed. Evaluating the specificity and applicability of these markers at the protein level to characterize Sertoli cells in culture would be critical. This review is expected to positively impact future work using primary cultures of rodent Sertoli cells.
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影响因子：
1.5
作者：
Bridges, Phillip J.;Koo, Yongburn;Ko, CheMyong
通讯作者：
Ko, CheMyong
影响因子：
2.5
作者：
AO, A;ERICKSON, RP;STALVEY, JRD
通讯作者：
STALVEY, JRD
影响因子：
4
作者：
Aslani, Ferial;Sebastian, Tim;Bhushan, Sudhanshu
通讯作者：
Bhushan, Sudhanshu
影响因子：
3.8
作者：
BEATTIE, PJ;WELSH, MJ;BRABEC, MJ
通讯作者：
BRABEC, MJ
DOI：
10.1186/1477-7827-1-4
发表时间：
2003-02-05
期刊：
Reproductive biology and endocrinology : RB&E
影响因子：
--
作者：
Baker, P J;Johnston, H;O'Shaughnessy, P J
通讯作者：
O'Shaughnessy, P J