Abstract
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.
Highlights
Spermatogonial stem cells (SSCs) are responsible for the producing of mature spermatozoa through a process of spermatogenesis[1]
Neonatal testes form 5-day-old piglets, PGP9.5-positive early spermatogonial cells were present in the centre of the seminiferous cord, and these cell in the luminal of seminiferous cord were translocated into basal compartment of seminiferous cords at p90
cluster of differentiation 14 (CD14)-expressing cells were located in the centre of the seminiferous cord, where PGP9.5-positive spermatogonial cells were found, in 5, 30, and 60-dayold testes (Fig. 1a–c) and were observed in PGP9.5-positive spermatogonia lining the basal lamina of seminiferous tubules in 90, 120, and 150-day-old testes (Fig. 1d–f)
Summary
Spermatogonial stem cells (SSCs) are responsible for the producing of mature spermatozoa through a process of spermatogenesis[1]. Integrins alpha-6 and beta-1, glial cell-derived neurotrophic factor receptor alpha 1 (GFRα1), cluster of differentiation 9 (CD9), CD90, and cadherin 1 (CDH1) are specific surface markers of SSCs in mouse testes[2,4,5,6]. These markers are useful for SSC isolation and the production of transgenic mice[7]. The surface expression of known SSC markers has not been well established in domestic animals, thereby limiting SSC studies. The potential use of CD14 as a surface marker of germ cells in porcine and its putative functions are discussed
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