组织工程与重建外科杂志

组织工程与重建外科杂志 >

2009 , Vol. 5 >Issue 1: 7 - 11

DOI: https://doi.org/10.3969/j.issn.1673-0364.2009.01.003

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大鼠睾丸间质细胞体外培养不同时间3β-羟基类固醇脱氢酶的表达

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  • 第二军医大学长海医院整形外科;上海交通大学医学院附属第九人民医院;上海组织工程重点实验室

网络出版日期: 2020-07-23

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Expression of 3β-Hydroxysteroid Dehydrogenase in Rat Leydig Cells Cultured by Differential Adhesion In Vitro

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  • 1 Department of Plastic Surgery;Changhai Hospital;Second Military Medical University;Shanghai 200433;China. 2 Shanghai Laboratory of Tissue Engineering;Shanghai Ninth People′s Hospital;Shanghai Jiaotong University School of Medicine;Shanghai 200011;China.;

Online published: 2020-07-23

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摘要

目的探讨通过差速贴壁法获得的7天龄、3周龄大鼠睾丸间质内细胞的细胞群体构成、体外培养不同时间3β-羟基类固醇脱氢酶(3β-HSD)的表达水平变化,以及细胞睾酮分泌功能变化。方法联合应用胶原酶消化、不锈钢滤网过滤及差速贴壁法获得7天龄、3周龄雄性Wistar大鼠睾丸间质组织内细胞,贴壁细胞以DMEM/F12培养液培养,分别对原代培养2h、4d细胞进行3β-HSD免疫化学染色及流式细胞分析,原代培养细胞同时给予人绒毛膜促性腺激素(HCG)刺激,测定HCG刺激组和非刺激组各代细胞培养液上清中睾酮水平及其对HCG刺激的反应。结果7天龄、3周龄鼠差速贴壁法获得的睾丸间质内细胞群经流式细胞鉴定,3β-HSD阳性细胞所占比例分别为(5.4±1.2)%、(59.2±3.2)%;培养4d后,两组3β-HSD阳性细胞比例分别为(93.6±1.2)%、(95.4±3.2)%。两组原代培养细胞均有睾酮生成功能,在HCG刺激下睾酮分泌均明显上升,7天龄组培养细胞睾酮分泌高峰迟于3周龄组。结论差速贴壁法获得的7天龄、3周龄大鼠睾丸间质细胞群中均含有部分Leydig干细胞(Stem Leydig cells,SLCs),SLCs在体外培养...

关键词： 睾丸间质细胞; 差速贴壁法; 睾酮; 3β-羟基类固醇脱氢酶; 组织工程

本文引用格式

毕宏达，王晓云，周广东，刘伟，李鸣，邢新 . 大鼠睾丸间质细胞体外培养不同时间3β-羟基类固醇脱氢酶的表达[J]. 组织工程与重建外科杂志, 2009 , 5(1) : 7 -11 . DOI: 10.3969/j.issn.1673-0364.2009.01.003

Abstract

Objective To investigate the constitution of cells population, changes in 3β-hydroxysteroid dehydrogenase (3β-HSD) expression and androgen productivity in rat Leydig cells cultured by differential adhesion during 4 days primary culture. Methods Testes from 7 days and 3 weeks old rats were harvested, then Leydig cells were isolated by using collagenase dispersion, stainless steel mesh infiltration and differential adhesion. 3β-HSD expression in cultured cells was observed by immunohistochemistry and flow Cyt...

Key words： Leydig Cell; Differential Adhesion; Testosterone; 3β-hydroxysteroid dehydrogenase; Tissue Engineering

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