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|本期目录/Table of Contents|

脐动脉平滑肌细胞体外培养方法的改进

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2014年第4期
页码:
425-428
栏目:
基础研究
出版日期:
2014-04-25

文章信息/Info

Title:
Improvement of in vitro culture method for umbilical artery smooth muscle cells
作者:
司静文1; 梁声茹2; 李军昌1; 王宗仁1
（第四军医大学第一附属医院：1.中医科，2.妇产科，陕西 西安 710032）
Author(s):
SI Jing-wen1;  LIANG Sheng-ru2;  LI Jun-chang1;  WANG Zong-ren1
(1.Department of Traditional Chinese Medicine, 2.Department of Gynecology and Obstetrics, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi, China)
关键词:
人脐动脉; 血管平滑肌细胞; 原代细胞培养
Keywords:
human umbilical artery;  vascular smooth muscle cell;  primary cell culture
分类号:
R329.2
DOI:
-
文献标识码:
A
摘要:
目的:建立并优化的人脐动脉血管平滑肌细胞(VSMCs)的体外培养方法。方法： 人脐动脉VSMCs的体外原代培养应用优化的组织块贴壁法进行。在分别应用传统细胞培养法和优化后细胞培养法贴壁培养提取的原代细胞10 d后，计数6孔板每孔细胞数目，并做统计比较分析相同时间内原代细胞的数量。以抗α-平滑肌肌动蛋白(α-SMA)抗体和抗CD31抗体进行免疫荧光染色和倒置相差显微镜观察鉴定培养的细胞。结果： 与传统细胞培养法相比，在相同的时间周期内优化实验方法后培养的原代细胞数量较传统多，我们可以在较短时间内得到实验所需的细胞量，因此优化后可缩短细胞的培养周期。原代和传代培养的脐动脉VSMCs生长良好，细胞经冻存、复苏后状态依然良好。光镜下细胞呈典型的“峰-谷”样生长，细胞多为长梭形，具有VSMCs的特征，且传代培养至第7代，细胞生长特性不变。经抗α-SMA、CD31抗体进行免疫荧光染色鉴定证实为VSMCs。结论： 优化的组织块贴壁法可缩短原代人脐动脉VSMCs的培养周期，且细胞的生物学特性较稳定，为与平滑肌相关的疾病进展及心血管生长发育的研究奠定了实验基础。
Abstract:
AIM:To establish and optimize the method for in vitro culture of human umbilical arterial smooth muscle cells (SMCs). METHODS: We counted each hole cell number in six orifices and statistically analyzed the number of primitive cells within the same time. Human umbilical artery vascular SMCs were cultured in vitro using optimized tissue-pieces inoculation. Alpha smooth muscle actin (alpha-SMA) and CD31 immunofluorescence staining methods and inverted phase contrast microscope were used for cell identification. RESULTS: Compared with the application of traditional cell culture technique, using the optimized cell cultivation this method to extract primitive cells was able to obtain more SMCs at the same time. The primary generation and subculture of SMCs grew well, and after frozen storage and recovery, the typical long spindle “peak-valley” growth of artery vascular SMCs could still be seen in the cultured cells. With subculture to the seventh generation, cell growth characteristics were stable and the cultured cells were confirmed as SMCs by alpha-SMA and CD31 immunofluorescence staining tests. CONCLUSION: Optimization of the tissue-pieces inoculation shortens the culture cycle time of human umbilical artery SMCs. Cultured cells have stable biological characteristics.

参考文献/References

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备注/Memo

备注/Memo:
收稿日期：2013-12-11.通讯作者：王宗仁，主任医师，主要从事中西医结合治疗心脏病基础和临床研究 Email：wangzongren@fmmu.edu.cn 作者简介：司静文，硕士生 Email：a15065895675@163.com
更新日期/Last Update: 2014-05-06