乳鼠软骨细胞体外培养及生物学特性研究
软骨细胞；,胶原酶；,原代培养,,软骨细胞；,胶原酶；,原代培养,1材料与方法,2结果,3讨论,参考文献：
摘要： 目的: 研究体外培养乳鼠胸骨剑突软骨细胞的原代培养方法，探讨该方法应用的可行性和应用价值。方法: 采用胶原酶消化法分离培养乳鼠胸骨剑突软骨细胞，以含20%胎牛血清的高糖DMEM培养液进行培养，并进行细胞接种与传代,用倒置显微镜、电镜及组织学方法观察细胞形态、超微结构。细胞采用ABC法进行Ⅱ型胶原免疫组化染色以及细胞形态观察鉴定软骨细胞起源。结果: (1)原代培养软骨细胞48 h后，大量细胞贴壁生长，呈圆形、长梭形或有2~3个突起，胞质透亮、饱满，7 d后细胞铺满整个培养皿底面。(2)原代软骨细胞呈圆形或多角形，第4代有一半变成梭形，第5代绝大部分变为长梭形；原代软骨细胞胞浆内粗面内质网和线粒体丰富，第3代软骨细胞生长增殖能力强于原代，第5代增殖能力明显减弱。结论: 体外培养的乳鼠剑突软骨细胞保持了其体内的基本特征，采用胶原酶消化法分离培养软骨细胞是一种可靠、简便、快速的细胞原代分离培养方法。第3代软骨细胞适合用于软骨组织工程及学术研究。关键词： 软骨细胞； 胶原酶； 原代培养
Culture in vitro and biological characteristics of newborn mouse chondrocytes
LI Liangzhong, ZHANG Donghui, XU Yonghua, et al
(Department of Labour Sanitary and Environment Sanitary, The Public Health College, Xinjiang Medical University, Urumqi 830000, China)
Abstract: Objective: To investigate the method of the primary culture chondrocyte from newborn mouse breastbone, and discuss the feasibility of this method and it's application value. Methods: We cutured chondrocyte which isolated by collagenase with 20% FBS and 80% high sugar DMEM and observed by inverted microscope, electron microscope and histology method. We identified the origination of chondrocyte by immunity histological chemistry of Ⅱ collagen and morphological observation. Results: (1) After 48h of primary culture, most chondrocytes grew adhering with the paste wall and showed round and long spindle shape or 2~3 processes, cytoplasm was translucent. After 7 days, chondrocytes spreaded whole dish surface. (2) The primary culture chondrocytes were round or polygonal shape, half of the 4th generation turned to spindleshape, most of the 6th passage were long spindleshape; The rough endoplasmic reticula and the mitochondrial were rich and welldeveloped, the ability of proliferation of the 3th generation were stronger than the first one, while the 5th generation was weaker. Conclusions: Primary culture chondrocyte from the newborn mouse breastbone maintains their basic characteristics. It is a reliable and simple method to isolate chondrocyte with collagenase in primary culture. The 3rd generation of chondrocyte are valiable of tissue engnieering and achedemic research. ......
您现在查看是摘要页，全文长 10042 字符。