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当前位置: 首页 > 文章 > 冷冻保存新生牛睾丸组织中支持细胞的分离纯化 中国兽医学报 2019 (1) 158-162
Position: Home > Articles > Separation and purification of Sertoli cells from cryopreserved testicular tissue of neonatal bulls Chinese Journal of Veterinary Science 2019 (1) 158-162

冷冻保存新生牛睾丸组织中支持细胞的分离纯化

作  者：
姜禹;赵欣欣;安星兰;蔡宁宁;张胜;王添;唐博;李子义;张学明
单  位：
吉林大学动物医学学院;吉林省添晟科技有限公司;吉林大学第一医院/转化医学研究院
关键词：
新生牛;睾丸组织;冷冻保存;支持细胞;差异贴壁
摘  要：
睾丸支持细胞(sertoli cells,SCs)是生精上皮中的多功能体细胞,其分离培养对体外探讨精子发生机理至关重要。因牛新鲜睾丸组织取材不便,本试验在前期基础上探讨了从冷冻保存的新生牛睾丸组织中分离纯化SCs的可行性。将冻存的1日龄牛睾丸组织复苏,进行形态学检查和SCs分离培养。石蜡切片苏木精-伊红(HE)染色显示,曲细精索及睾丸间质保存完好。二步酶消化结果显示,生精上皮解离良好。利用差异贴壁法将原代细胞纯化后扩大培养,绝大多数细胞呈不规则多边形,3d即可生长至汇合。RT-PCR分析显示,贴壁细胞表达SCs阳性分子标记胶质细胞源神经营养因子(GDNF)和雄性激素结合蛋白(ABP)而不表达精原细胞分子标记PLZF。流式细胞术检测显示,所获细胞中ABP和GDNF阳性细胞数分别为99.7%和99.8%。以上结果表明,从冻存的新生牛睾丸组织中可获高纯度的SCs。本试验结果为后续相关试验奠定了基础。
译  名：
Separation and purification of Sertoli cells from cryopreserved testicular tissue of neonatal bulls
作  者：
JIANG Yu;ZHAO Xin-xin;AN Xing-lan;CAI Ning-ning;ZHANG Sheng;WANG Tian;TANG Bo;LI Zi-yi;ZHANG Xue-ming;College of Veterinary Medicine,Jilin University;Academy of Translational Medicine,First Hospital,Jilin University;Jilin Tiansheng Technology Co.,Ltd;
单  位：
JIANG Yu%ZHAO Xin-xin%AN Xing-lan%CAI Ning-ning%ZHANG Sheng%WANG Tian%TANG Bo%LI Zi-yi%ZHANG Xue-ming%College of Veterinary Medicine,Jilin University%Academy of Translational Medicine,First Hospital,Jilin University%Jilin Tiansheng Technology Co.,Ltd
关键词：
neonatal bull;;testicular tissue;;cryopreservation;;sertoli cell;;differential plating
摘  要：
Sertoli cells(SCs)are multi-functional somatic cells in the seminiferous epithelium.The isolation and in vitro culture of SCs are of great importance to probe the spermatogenesis mechanism.Since it is inconvenient to get bovine testicular tissues,here we investigated the possibility of SCs isolation and enrichment from cryopreserved neonatal bovine testis based on our previous work.The cryopreserved testicular tissues of postnatal 1-day-old bulls were thawed for morphological examination and SCs separation.Hematoxylin-eosin(HE)staining of the paraffin sections showed that the seminiferous cords and interstitium were well preserved.The two-step enzymatic digestion showed that the seminiferous epithelia were dissociated completely.The attached primary cells were enriched by differential plating,and expanded subsequently.Most of them were in irregular polygon shape and grew to confluence within 3days.RT-PCR analysis revealed that they expressed SCs marker GDNF and ABP but not spermatogonial marker PLZF.Flow cytometry analysis showed that the enriched cells contained high purified ABP+cells(99.7%)and GDNF+cells(99.8%).The above data indicate that high purified SCs can be obtained from cryopreserved testicular tissues of neonatal bulls,which lays a solid foundation for our next step.

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