山羊精原干细胞的分离纯化及鉴定

doi:10.11843/j.issn.0366-6964.2013.10.007

Abstract

Abstract:

The present study was aimed to establish a novel method that can efficiently isolate and purify spermatogonial stem cells (SSCs) from goat testis. Single cell suspension was prepared from testis of 4-month old goat by three-step enzymatic digestion. The SSCs were enriched from the suspended cells via differential attachment in gelatin, collagen, and laminin coated dishes respectively, according to the differential adherence capability in different kinds of cells. Immunofluorescent staining by using SSC molecular marker PLZF and CDH1 was performed for analysis of enrichment efficiency of the SSCs. The percentages of SSCs were calculated through PLZF- and CDH1- positive cells. The enriched cells were primarily cultured and carried out identification. 1.36×10⁷ suspended cells were harvest from per gram of goat testis. 8.7×10⁴ cells were obtained after purification, in which the purity of SSCs was 87.0%. The purified cells could be cultured to form clusters in vitro, and CDH1 and PLZF were identified to express in the cells by immunofluorescent staining. The results show that a technique for purification of goat SSCs has been established by three-step enzymatic digestion and differential attachment. Goat SSCs purified by this technique can be cultured in vitro for proliferation and forming clusters.

CLC Number:

S827
Q28

KONG Qun-fang, LUO Fen-hua, WU Sachula, BAO Jia-jing, LIU Dai-yan, WU Ying-ji. Enrichment and Identification of Spermatogonial Stem Cells from Goat Testis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(10): 1554-1560.

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Enrichment and Identification of Spermatogonial Stem Cells from Goat Testis

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