The molecular signature and spermatogenesis potential of newborn chicken spermatogonial stem cells in vitro

Abstract

Although chicken spermatogonial stem cells (SSCs) have received considerable attention in recent years, only a few studies so far have focused on their derivation and characterization in vitro. Identification of specific molecular biomarkers and differentiation capacity of chicken SSCs would not only help us to understand cell and molecular biology of these cells, but also can contribute to their applications in biotechnology. In this regard, we found that colony-forming cells (SSCs) in newborn chicken testicular cell cultures were positive for alkaline phosphatase activity and also expressed specific markers including DAZL, STRA-8, CVH, PLZF, SPRY-1, GFRα1, GDNF, POU5F1, NANOG, GPR125, THY-1, c-KIT, and BCL6B, at mRNA level. Moreover, these cells expressed POU5F1 and GPR125 proteins as reliable intracellular and cell surface markers, respectively; whereas they were negative for SSEA-1. Furthermore, we showed that newborn chicken colony-forming cells had spermatogenesis potential and thus could be produced sperm-like cells in a three-dimensional matrix in vitro. In conclusion, this study reports novel insights into the molecular signature of newborn chicken SSCs in comparison with mammalian SSCs and for the first time we report a successful protocol for in vitro spermatogenesis and thus production of sperm-like cells from newborn chicken testicular cell cultures.

Isolation and Culture of Pig Spermatogonial Stem Cells and Their in Vitro Differentiation into Neuron-Like Cells and Adipocytes

“…Their rarity has presented problems in isolation and culturing them and the development of successful techniques for doing so has been the focus of intense study. At present, SSCs have been successfully isolated and cultured from many species, including human, mice [ 5 ], cattle [ 6 ], pig [ 7 , 8 , 9 ], goat [ 10 , 11 , 12 ], dog [ 13 ], chicken [ 14 , 15 ], and even fish [ 16 ].…”

Biotechnological Potential of Chicken Stem Cells

“…Alternative strategies for obtaining SSCs have also been introduced such as differentiation of MSCs to SSC-like cells [7,9,20]. Recently, we successfully could isolate and maintain SSCs in laboratory conditions and could determine their molecular signatures and their differentiation potential [21,22]. Understanding the stem cell differentiation mechanisms and their maintenance in vitro will help to the scientific community to produce transgenic animals by manipulating stem cells easily with a great biotechnology potential.…”

ISOLATION AND CHARACTERIZATION OF ROOSTER (Gallus gallus) SPERMATOGONIA

“…Isolation and characterization of spermatogonial cells are reported for laboratory and farm animals, such as mice [13,14] and hamsters [15], pigs [16][17][18][19][20] and goat [21,22], and in fishes [23]. The spermagonial cells were also isolated from the gonads of embryos and testes of day-old chicks [24,25].…”