湖北白猪精原干细胞分离培养及初步鉴定

doi:10.14088/j.cnki.issn0439-8114.2021.22.027

湖北农业科学 ›› 2021, Vol. 60 ›› Issue (22): 128-132.doi: 10.14088/j.cnki.issn0439-8114.2021.22.027

湖北白猪精原干细胞分离培养及初步鉴定

张立苹, 华再东, 肖红卫, 任红艳, 朱喆, 毕延震

湖北省农业科学院畜牧兽医研究所/动物胚胎工程及分子育种湖北省重点实验室,武汉 430064

收稿日期:2021-08-13 出版日期:2021-11-25 发布日期:2021-12-10
作者简介:张立苹（1982-）,女,河北邢台人,助理研究员,硕士,研究方向为动物胚胎工程,（电话）13477076406（电子信箱）949164559@qq.com。
基金资助:
湖北省农业科技创新中心项目（2019-620-000-001-20）; 湖北省农业科学院院青年拔尖人才项目（Q2018020）; 湖北省农业科学院领军人才项目（L2018015）; 湖北省自然科学基金创新群体项目（2018CFA014）; 湖北省技术创新重大专项（2019ABA084）; 国家自然科学基金项目（31772577）; 湖北省自然科学基金杰出青年项目（2018CFA043）; 湖南创新型省份建设专项经费（2019RS1068）

Isolation, culture and identification of Hubei porcine spermatogonial stem cells

ZHANG Li-ping, HUA Zai-dong, XIAO Hong-wei, REN Hong-yan, ZHU Zhe, BI Yan-zhen

Hubei Key Laboratory of Animal Embryo Engineering and Molecular Breeding,Institute of Animal Science and Veterinary Medicine,Hubei Academy of Agricultural Sciences,Wuhan 430064,China

Received:2021-08-13 Online:2021-11-25 Published:2021-12-10

摘要/Abstract

摘要： 以5日龄、10日龄和15日龄湖北白猪睾丸为主要研究对象,采用浓度为4 mg/mL Ⅳ型胶原蛋白酶联合0.25%胰蛋白酶两步酶法消化法获取湖北白猪精原干细胞,通过低速离心和差异贴壁法对湖北白猪精原干细胞进行纯化,以湖北白猪睾丸支持细胞作为饲养层进行体外培养,结合碱性磷酸酶（AKP）进行染色鉴定。结果表明,培养获得的细胞即为目的细胞SSCs。

关键词: 湖北白猪, 精原干细胞, 睾丸, 碱性磷酸酶

Abstract: The present study amimed to estabish a ripe system of Hubei white pig SSCs in vitro isolation and culture, in order to lay the foundation for related research. The testes of Hubei white pigs aged 5days, 10 days and 15 days after birth were studied,to obtained the spermatogonial stem cells of Hubei white pig by two-step enzyme digestion（4 mg/mL type Ⅳ collagenase and 0.25% trypsin）. The spermatogonial stem cells of Hubei white pigs were purified by low-speed centrifugation and differential adherence method and cultured in the sertoil cells of Hubei white pig that as feeder layer in vitro. Identified by alkaline phosphatase （AKP） and the results showed that the cultured cells were SSCs.

Key words: Hubei pig, spermatogonial stem cells, testis, alkaline phosphatase

中图分类号:

Q813.1⁺1

张立苹, 华再东, 肖红卫, 任红艳, 朱喆, 毕延震. 湖北白猪精原干细胞分离培养及初步鉴定[J]. 湖北农业科学, 2021, 60(22): 128-132.

ZHANG Li-ping, HUA Zai-dong, XIAO Hong-wei, REN Hong-yan, ZHU Zhe, BI Yan-zhen. Isolation, culture and identification of Hubei porcine spermatogonial stem cells[J]. HUBEI AGRICULTURAL SCIENCES, 2021, 60(22): 128-132.

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湖北白猪精原干细胞分离培养及初步鉴定

Isolation, culture and identification of Hubei porcine spermatogonial stem cells

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