摘要
目的探讨从兔肋软骨膜和肋软骨中分别提取间充质干细胞的方法以及两者间的比较。方法取3～4周新西兰大白兔,共20只。酶消化及贴壁培养法分别获取软骨膜细胞和软骨细胞,体外培养观察其生长、形态、表型特征及其诱导成骨、成脂的分化潜能。结果从肋软骨膜中分离的原代细胞聚集成团,多为圆形和多边形;从第2代开始细胞以梭形为主,呈漩涡状排列,已传至第15代。从肋软骨中分离的原代细胞连接成片,以铺路石形态为主,第2代和第3代细胞逐渐转变成梭形;但第5代出现细胞老化,只传至第6代。免疫细胞化学染色发现,两种原代细胞均不表达CD29、CD90和CD34,从第2代开始均表达CD29、CD90,仍不表达CD34。原代软骨细胞表达Ⅱ型胶原,第2代细胞弱表达,第3代细胞开始不表达Ⅱ型胶原;而软骨膜细胞不表达Ⅱ型胶原。两种来源的第3代细胞经诱导均可分化为成骨细胞或者脂肪细胞。结论从兔肋软骨膜和软骨中均可获得干细胞。软骨细胞在体外可去分化成为干细胞。但软骨膜来源的干细胞的生长明显优于软骨来源的干细胞。
Objective To separate rabbit-rib perichondrium cells and cartilage cell, and compare their characteristics. Methods Rib perichondrium cells and rid cartilage ceils were isolated from New Zealand white rabbits, aged 3 and 4 weeks by the enzyme digest and adherent culture method and then cultured in vitro. The growth, phenotype and potention of dipogenic and osteogenic differentiation in both types of the cells were observed. Results In the primary generation of rib perichondrium cells were round or polygon and assemble as conglobation; The cells from two-passage began to show a spindle-shape and whirlpool arrangement; and the cells grew to the 15 th generation. The primary generation of rib cartilage cells was in cobblestone appearance, and then gradually changed as spindle-shape cells from two- passage to there-passagee cells. But the cells appeared senescence in the five-passage, and then were dead gradually. The results of immunocytochemical stain showed that CD29, CD90 and CD34 were negative in primary rid perichondrium and cartilage cells. From the two passage cells of rib perichondrium and rid cartilage began expressing CD29 and CD90, but not CD34. The collogen type 11 was expressed by the primary rid cartilage cells, and then vanishied in the three-passage cells. The adipogenic and osteogenic differentiation were induced in three-passage rib perichondrium and rid cartilage cells in vitro. ConcLusion The stem cells can be isolated from rib perichondrium and cartilage of rabbit. The cartilage cell may become the stem cell through dedifferentiation in vitro. The growth of the stem cell derived from rib perichondrium was surpassed more than that derived from rib cartilage. [
出处
《解剖学报》
CAS
CSCD
北大核心
2012年第5期641-646,共6页
Acta Anatomica Sinica
关键词
肋软骨膜
肋软骨
干细胞
生物学特性
免疫组织化学
兔
Rib perichondrium
Rib cartilage
Stem cell
Biological characteristic
Immunohistochemistry
Rabbit
