Abstract
Gonocytes are progenitors of spermatogonial stem cells in the neonatal testis. We have previously shown that upon culturing, neonatal porcine gonocytes and their colonies express germ cell and pluripotency markers. The objectives of present study were to investigate in vitro trans-differentiation potential of porcine gonocytes and their colonies into cells from three germinal layers, and to assess pluripotency of cultured gonocytes/colonies in vivo. For osteogenic and tri-lineage differentiation, cells were incubated in regular culture media for 14 and 28 days, respectively. Cells were cultured for an additional 14 days for osteogenic differentiation or 7 days for differentiation into derivates of the three germinal layers. Osteogenic differentiation of cells and colonies was verified by Alizarin Red S staining and tri-lineage differentiation was confirmed using immunofluorescence and gene expression analyses. Furthermore, upon implantation into recipient mice, the cultured cells/colonies developed teratomas expressing markers of all three germinal layers. Successful osteogenic differentiation from porcine germ cells has important implications for bone regeneration and matrix formation studies. Hence, gonocytes emerge as a promising source of adult pluripotent stem cells due to the ability to differentiate into all germinal layers without typical biosafety risks associated with viral vectors or ethical implications.
Highlights
Gonocytes are a transitory population of male germline stem cells (MGSCs) in the neonatal testis; they develop from fetal primordial germ cells (PGCs) and postnatally transition into spermatogonial stem cells (SSCs)
We showed that implantation of cultured gonocytes and their colonies into the subcutaneous tissue of recipient mice can lead to development of tumor-like growth formations
Neonatal porcine gonocytes represent a promising source of adult stem cells
Summary
Gonocytes are a transitory population of male germline stem cells (MGSCs) in the neonatal testis; they develop from fetal primordial germ cells (PGCs) and postnatally transition into spermatogonial stem cells (SSCs). SSCs, considered adult stem cells, are fundamental for spermatogenesis due to their dual potential for self-renewal and differentiation ability for subsequent mitosis and meiosis [4,5] Compared to both PGCs and SSCs, gonocytes in the neonatal testis are a preferred source of MGSCs, since they offer a relatively large population, ease of accessibility, identifiable morphology, and more specific molecular markers for their identification and isolation [6]. Compiling evidence demonstrates the dedifferentiation potential of MGSCs, as well as their ability to derive multipotent or even pluripotent stem cells in an ex situ environment [9,10,11,12,13,14] These cells can spontaneously transform into a population of cells that express embryonic stem (ES) cell markers, otherwise if genetic manipulations were needed, they would have imposed safety concerns regarding their use and expansion [9,15,16,17]. The ability to dedifferentiate into a pluripotent state allows these ES-like cells to convert into other cell types from derivatives of three germinal layers [9,10,11,13]
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