Department of Stem Cell and Regenerative Technology, KIT, Konkuk University, 120 Neungdongro, Gwangjin-gu, Seoul, 05029, Republic of Korea.
Department of Beef Science, Korea National College of Agricultures and Fisheries, Jeonju-si, Jeonbuk, 54874, Republic of Korea.
Sci Rep. 2019 Jul 10;9(1):9980. doi: 10.1038/s41598-019-46000-6.
Molecular markers of spermatogonia are necessary for studies on spermatogonial stem cells (SSCs) and improving our understanding of molecular and cellular biology of spermatogenesis. Although studies of germ cell surface marker have been extensively conducted in the testes of rodents, these markers have not been well studied in domestic animals. We aimed to determine the expression pattern of cluster of differentiation 14 (CD14) in developing porcine testes and cultured porcine SSCs (pSSCs), as well as its role in pSSC colony formation. Interestingly, expression of CD14 was observed in porcine testes with PGP9.5-positive undifferentiated spermatogonia at all developmental stages. In addition, in vitro cultured pSSCs expressed CD14 and showed successful colony formation, as determined by fluorescence-activated cell sorting and flow cytometry. PKH26 dye-stained CD14-positive cells transplants were performed into the testes of recipient mice, which were depleted of both testicular germ and somatic cells from immunodeficiency mice and were shown to colonise the recipient testes. Moreover, a colony-forming assay showed that the development of pSSC colonies was disrupted by a high concentration of lipopolysaccharide. These studies indicated that CD14 is surface marker of early spermatogonia in developing porcine testes and in pSSCs, suggesting a role for CD14 in porcine spermatogenesis.
精原干细胞（SSCs）的分子标记对于研究 SSCs 和提高我们对精子发生的分子和细胞生物学的理解是必要的。虽然在啮齿动物的睾丸中已经广泛研究了生殖细胞表面标记物，但这些标记物在家畜中尚未得到很好的研究。我们旨在确定 CD14 在发育中的猪睾丸和培养的猪 SSCs（pSSCs）中的表达模式，以及其在 pSSC 集落形成中的作用。有趣的是，在所有发育阶段，PGP9.5 阳性未分化精原细胞的猪睾丸中都观察到 CD14 的表达。此外，通过荧光激活细胞分选和流式细胞术确定，体外培养的 pSSC 表达 CD14 并成功形成集落。将 PKH26 染料染色的 CD14 阳性细胞移植到受体小鼠的睾丸中，这些受体小鼠的睾丸生殖和体细胞均从免疫缺陷小鼠中耗尽，并显示出能够定植受体睾丸。此外，集落形成测定表明，高浓度的脂多糖破坏了 pSSC 集落的发育。这些研究表明，CD14 是发育中的猪睾丸和 pSSCs 中早期精原细胞的表面标记物，表明 CD14 在猪精子发生中起作用。
