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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۲۱-۲۱
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عنوان انگلیسی Ps-1: Comparison of Differential Plating and MACS in Enrichment of Goat Undifferentiated Spermatogonia
چکیده انگلیسی مقاله Objective: Spermatogonia are male germ line progenitors of sperm cells. In bovine, rodents and primates, undifferentiated spermatogonia and spermatogonial stem cells (SSCs) express the surface marker THY1. Since cells can be enriched from mixed populations by positive or negative selection on physical or functional characteristics such as cell surface markers, present study was aimed to compare the efficiency of two different enrichment methods: Magnetic activated cell sorting (MACS) and differential plating (DP) in pre-pubertal goat testes cell population. To comparison of two enrichment methods efficiency, real time PCR analysis, protein expression, and transplantation into recipient mice data were analyzed. For control group, the intact testis cells population were used. Materials and Methods: After 3 times enzymatic digestion of pre-pubertal goat testicular tissues, cells were cultured for 16 h in laminin-coated flasks and DP carried out. For MACS method, cell suspension were sorted using THY1 surface marker. Results: Real time PCR analysis revealed that expression of THY1, PLZF, VASA, BCL6B, UCHL1 as SCCs characteristic genes in THY1 positive cells were significantly higher than in the DP isolation cells and unselected total cells. Transplantation of THY1positive, DP isolated and intact population of testis cells (control) into the mice recipient seminiferous tubules revealed that THY1 positive cells had higher capacity for colony formation were compared to DP isolated and intact cells. Conclusion: The results indicated that isolation of undifferentiated testis cells using MACS method can be more efficient than differential plating.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/807
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