摘要
目的探讨先天性小耳畸形残耳软骨细胞体外生长的生物学特性。方法取先天性小耳畸形患者外耳再造手术中废弃的残耳软骨组织，采用胶原酶消化法获得残耳软骨细胞；体外传代培养及生物学特性研究：细胞形态学观察，细胞生长曲线测定，Ⅱ型胶原免疫组化鉴定残耳软骨细胞，RT—PCR检测第1～5代残耳软骨细胞的Ⅱ型胶原及蛋白多糖的mRNA表达。结果原代和第1-3代残耳软骨细胞生长旺盛，呈多角形，第3代以后细胞体积变大，呈长梭形转化，细胞增殖逐渐减弱；免疫组化检测第2代残耳软骨细胞Ⅱ型胶原表达呈阳性；RT—PCR显示mRNA水平第1～3代软骨细胞Ⅱ型胶原高表达，随传代次数增加表达逐渐减弱，第5代基本消失；第1～3代细胞蛋自多糖呈高表达，第4代以后随传代表达逐渐减弱。结论体外成功分离培养出残耳软骨细胞，第1—3代细胞形态保持稳定．Ⅱ型胶原和蛋白聚糖稳定旱高表达。
Objective To investigate the bionomics of chondrocytes from congenital microtia in vitro. Methods The cartilaginous tissue from congenital microtia were collected and the chondrocytes were isolated by the method of collagenase digestion, the isolated cells were cultured with serial subcnhivation and then the cultured cells were supplied for the research of bionomics ： Morphological changes observation, cell growth curve measurement, chondrocytes identification with immunohistochemisty which collagen type Ⅱ was as monoclonal antibody and the mRNA expression of proteoglyeans and collagen type Ⅱ from 1- 5 passages ehondrocytes de- tection with RT-PCR method. Results The cells from primary and 1 -3 passages were eugenic and showed polygon; the cell volume was bigger; the shape was turned into spindle-shape and cell proliferation increased gradually after the 3rd passage; the expression of collagen type Ⅱ was positive by immunohistochemisty at the 2nd passage; the mRNA expression of collagen type Ⅱ from 1 - 3 passages was stronger which decreased gradually with passage times and was negative at the 5th passage; the expression of proteoglycans from 1 -3 passages was stronger and decreased gradually after the 4th passages. Conclusion The residual auricular chondrocytes are successfully isolated and cultured in vitro. Cell morphology from 1 st to 3rd passage remained stable, and the expression of Ⅱ collagen and proteoglycans is stronger and stable.
出处
《中国美容整形外科杂志》
CAS
2010年第9期553-556,共4页
Chinese Journal of Aesthetic and Plastic Surgery
关键词
残耳软骨细胞
Ⅱ型胶原
蛋白多糖
种子细胞
软骨组织工程
Residual auricular chondrocytes
Collagen type Ⅱ
Proteoglycans
Seed cell
Cartilage tissue engineering
