THY1 as a reliable marker for enrichment of undifferentiated spermatogonia in the goat

被引:54
作者
Abbasi, Hassan [1 ,2 ]
Tahmoorespur, Mojtaba [1 ]
Hosseini, Sayyed Morteza [2 ]
Nasiri, Zahra [2 ]
Bahadorani, Mehrnoosh [2 ,3 ]
Hajian, Mahdi [2 ]
Nasiri, Mohammad Reza [1 ]
Nasr-Esfahani, Mohammad Hossein [2 ]
机构
[1] Ferdowsi Univ Mashhad, Fac Agr, Dept Anim Sci, Mashhad, Iran
[2] ACECR, Royan Inst Biotechnol, Reprod Biomed Res Ctr, Dept Reprod Biotechnol, Esfahan, Iran
[3] Islamic Azad Univ, Dept Biol, Falavarjan Branch, Esfahan, Iran
关键词
Undifferentiated spermatogonia; Goat; Testis; THY1; MACS; CELL SELF-RENEWAL; STEM-CELLS; FUNCTIONAL-CHARACTERISTICS; NUCLEAR TRANSFER; GENE-EXPRESSION; TRANSPLANTATION; CULTURE; IDENTIFICATION; VASA; CRYOPRESERVATION;
D O I
10.1016/j.theriogenology.2013.07.020
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Spermatogonial stem cells are unique cells of testes that can restore fertility upon transplantation into recipient testes. However, use of suitable markers for enrichment of these cells have important potential application. THY!, is an established conserved marker of spermatogonial stem cells in bovine, rodents, and primates, but there is no information available in goats. After three rounds of enzymatic digestion of prepubertal goat testicular tissues, undifferentiated spermatogonia positive for THY1 were isolated by magnetic-activated cell sorting and were used for immunocytochemistry, real-time polymerase chain reaction analysis for gene expression, protein expression, and transplantation into recipient mice. Immunocytochemical analyses showed that significantly higher percentage of THY1(+) cells were positive for PLZF and VASA when compared with unselected population. This result for PLZF was further confirmed at the protein level. Real-time polymerase chain reaction analysis revealed that expression of THY1, PLZF, VASA, BCL6B, and UCHL1 as SCCs characteristic genes in THY1(+) cells was significantly higher than in the initial population. Finally, transplantation of PKH26-labeled cells revealed that THY1(+) cells had higher capacity for colony formation when compared with unselected cells. In conclusion, the results provide indications that THY1 surface marker can be reliably used for enrichment of undifferentiated spermatogonial in the goats. (C) 2013 Elsevier Inc. All rights reserved.
引用
收藏
页码:923 / 932
页数:10
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