中国组织工程研究Issue(11)：1901-1908,8.DOI:10.3969/j.issn.2095-4344.2013.11.001
不同代次兔髓核细胞的形态学特征和生物学性状*★
Different generations of rabbit nucleus pulposus cells:Morphological and biological properties
摘要
Abstract
BACKGROUND: Intervertebral disc degeneration is a chronic and complicate procedure, but its mechanism is stil unclear and difficult for self-repairs. Recent studies addressing stem cel transplantation in the treatment of disc degeneration disease are stil limited to the laboratory study. Research on biological properties of nucleus pulposus cells can provide theoretical basis for studying the mechanism underlying intervertebral disc degeneration, construction of the intervertebral disc by tissue engineering, and gene therapy. OBJECTIVE: To investigate the characteristics of different generations of rabbit nucleus pulposus cells, searching for the best suitable seed cells to treat degenerative disc diseases. METHODS: Nucleus pulposus cells from New Zealand white rabbits were separated, cultured and then passaged. The morphological changes of primary, passages 3 and 4 nucleus pulposus cells were observed by hematoxylin-eosin staining under an inverted microscope. The biological properties of rabbit nucleus pulposus cells were observed. Aggrecan and type Ⅱ colagen expressions w ere detected by toluidine blue and immunocytochemistry staining, respectively. Type nucleus pulposus cells were detected by reverse transcription-PCR. RESULTS AND CONCLUSION: Rabbit nucleus pulposus cells were successful y cultured and passaged in vitro. Primary nucleus pulposus cells were round or polygonal, and the average adherence time was 7 days. The first and third generations of nucleus pulposus cells were round or polygonal, and have strong vitality. Hematoxylin-eosin staining showed that nuclei were in a uniform blue-black, and cytoplasm showed light pink. The cytoplasm of nucleus pulposus cells was sky blue stained for toluidine blue staining, and type II col agen immunohistochemical staining showed the cytoplasm of nucleus pulposus cells displayed yel owish-brown. Passage 4 nucleus pulposus cells appeared with degeneration, and type mRNA expression was significantly decreased compared with previous generations. The first three generations of nucleus pulposus cells were exuberant in metabolism and showed consistent phenotypes and normal expression of aggrecan and type Ⅱ col agen. Passage 4 nucleus pulposus cells began to age and degenerate.关键词
组织构建/软骨组织构建/兔/髓核细胞/体外培养/聚集蛋白聚糖/Ⅱ型胶原/椎间盘退变/细胞培养/省级基金/组织构建图片文章Key words
tissue construction/cartilage tissue construction/rabbits/nucleus pulposus cells/in vitro culture/aggrecan/type Ⅱ col agen/intervertebral disc degeneration/cel culture/provincial grants-supported paper/tissue construction photographs-containing paper分类
医药卫生引用本文复制引用
武海军,银和平,李树文,白明,杜志才,曹振华..不同代次兔髓核细胞的形态学特征和生物学性状*★[J].中国组织工程研究,2013,(11):1901-1908,8.基金项目
内蒙古自治区自然科学基金项目资助(2009MS1117)。Supported by：the Natural Science Foundation of Inner Mongolia Autonomous Region, No.2009MS1117* （2009MS1117）
