驴皮成纤维细胞的体外培养研究

doi:10.16431/j.cnki.1671-7236.2017.08.006

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (8): 2255-2260.doi: 10.16431/j.cnki.1671-7236.2017.08.006

驴皮成纤维细胞的体外培养研究

王艳萍^1,2, 高帅³, 刘迥¹, 曾维斌^1,2

1. 石河子大学动物科技学院, 石河子 832003;
2. 塔里木大学动物科学学院, 新疆生产建设兵团塔里木畜牧科技重点实验室, 阿拉尔 843300;
3. 中国农业大学动物科技学院, 北京 100193

收稿日期:2017-03-21 出版日期:2017-08-20 发布日期:2017-08-18
通讯作者: 曾维斌 E-mail:zwbdky@126.com
作者简介:王艳萍(1978-),女,甘肃渭源人,硕士,副教授,研究方向:基础兽医学和动物生殖技术,E-mail:wypdky@126.com
基金资助:
国家自然科学基金项目（31260531、31360530）；兵团塔里木畜牧科技重点实验室项目（HS201503）

Study on in vitro Culture of Donkey Skin Fibroblasts

WANG Yan-ping^1,2, GAO Shuai³, LIU Jiong¹, ZENG Wei-bin^1,2

1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;
2. Key Laboratory of Tarim Animal Husbandry Science & Technology of Xinjiang Production and Construction Groups, College of Animal Science, Tarim University, Alar 843300, China;
3. College of Animal Science and Technology, China Agricultural University, Beijing 100193, China

Received:2017-03-21 Online:2017-08-20 Published:2017-08-18

摘要/Abstract

摘要：

试验旨在比较不同培养方法对驴皮成纤维细胞培养的效果，以便建立其体外高效快速的培养体系。采集6~7岁健康的新疆驴真皮组织，分别用组织块贴壁法和双酶消化法对驴皮成纤维细胞进行原代培养，并检测分析细胞的生长特点、生长速度、细胞周期与支原体污染等指标。结果显示，双酶消化法（0.25%胰酶处理1 h，再用0.5%胶原蛋白酶Ⅰ处理6 h）培养驴皮组织3 d后，成纤维细胞进入了对数增殖期，而组织块贴壁法则需要15 d；细胞周期经流式分析发现，处于G0/G1期与S+G2+M期的细胞均约为50%，其余5.17%细胞处于凋亡期，表明大多细胞处于分裂增殖的活跃期，细胞具有很强的活力；试验中培养的细胞均无支原体污染。综上，应用双酶消化法可快速、高效地建立驴皮成纤维细胞体外培养体系。

关键词: 驴皮; 成纤维细胞; 组织块贴壁法; 双酶消化法

Abstract:

This study was aimed to compare different culture methods for donkey skin fibroblasts to establish a highly efficient and rapid culture system. The dermal tissue of the healthy and 6-7 year old Xinjiang donkey was cultured for primary cells of skin fibroblast with tissue explants adherent method and double enzyme digestion method. Then,the growth characteristics,proliferation,cell cycle and mycoplasma contamination were detected and analyzed. The results showed that the fibroblast cells entered the logarithmic growth phase after cultured for 3 days by double enzyme digestion(0.25% trypsin digestion for 1 h and then 0.5% collagenase Ⅰ for 6 h), while it required 15 days with tissue explants adherent method. Flow cytometry analysis showed that nearly half of the cells were in G0/G1 stage,the other half in S+G2+M stage, while just 5.17% of the cells were apoptosis,which indicated that most of the cells were in the active stage of division and proliferation,and had strong vitality. Moreover, there was no mycoplasma contamination in the cultured cells. In summary,the culture system of donkey skin fibroblasts was rapidly and effectively established with the double enzyme digestion method in vitro.

Key words: donkey skin; fibroblasts; tissue explants adherent method; double enzyme digestion method

中图分类号:

Q813.1

王艳萍, 高帅, 刘迥, 曾维斌. 驴皮成纤维细胞的体外培养研究[J]. 《中国畜牧兽医》, 2017, 44(8): 2255-2260.

WANG Yan-ping, GAO Shuai, LIU Jiong, ZENG Wei-bin. Study on in vitro Culture of Donkey Skin Fibroblasts[J]. , 2017, 44(8): 2255-2260.

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驴皮成纤维细胞的体外培养研究

Study on in vitro Culture of Donkey Skin Fibroblasts

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