中国现代医学杂志2016，Vol.26Issue(18)：1-5,5.DOI:10.3969/j.issn.1005-8982.2016.18.001
改良分次酶消化法分离培养兔肌腱干细胞及诱导分化鉴定*
Isolation and culture of rabbit tendon stem cells by modified fractional enzyme digestion and identification of their induced differentiation
摘要
Abstract
Objective To isolate and culture rabbit tendon stem cells in vitro by a modified fractional enzyme digestion method, observe their biological characteristics, and induce and identify their differentiation. Methods Rabbit patellar tendon tissue was removed under sterile condition. After the tissue was digested by a modified fractional enzyme digestion method and enzyme digestion method respectively, a low-density dilution inoculation method was adopted to isolate, culture and passage the rabbit tendon stem cells. Morphological characteristics of the cells were observed under an inverted phase contrast microscope and the growth curves of the cells isolated from the tendon tissue with the two methods were drawn. The expressions of tendon stem cell surface antigen markers were identified by a flow cytometer. The P3-P4 generations of the tendon stem cells were induced for the differentiation into osteoblasts and chondroblasts, and the differentiation was identified. Results The proliferation of tendon stem cells isolated by the modified fractional enzymatic digestion and inoculated with low-density dilution inoculation method was accelerated, and the cell shapes were uniform with few impure cells. Surface antigen markers CD90 and CD44 of the isolated tendon stem cells were positive, but CD34 and CD14 were negative, indicating that the isolated cells were the tendon stem cells and the identified tendon stem cells had the ability of differentiation into osteoblasts and chondroblasts. Conclusions The modified fractional enzyme digestion for isolation and culture of rabbit tendon stem cells is simple and practicable. The growth and passage speed of the cells are satisfactory, their viability is good, and the purity is high. In addition, the successful isolation and culture of tendon stem cells may create a new way for the research on tendon-related diseases.关键词
改良酶消化法/肌腱干细胞/分化鉴定Key words
modified enzyme digestion method/tendon stem cell/differentiation and identification分类
医药卫生引用本文复制引用
张立岩,柴伟,孙新,田丹,徐睿,雷昊,艾津辉,王华,王海涛,陈继营..改良分次酶消化法分离培养兔肌腱干细胞及诱导分化鉴定*[J].中国现代医学杂志,2016,26(18):1-5,5.基金项目
国家自然科学基金委员会资助项目（81301564）；吉林省教育厅“十二五”科学技术研究项目（吉教科合字【2015】第141号）；吉林省科技厅项目（20130624003JC）；吉林省教育厅“十二五”科学技术研究项目（吉教科合字【2011】第132号）；吉林市科技项目 （）
