人胎关节软骨细胞体外培养的生物学特性

The biological characteristics of human fetal articular chondrocytes cultured in vitro

目的:研究软骨组织工程中传代软骨细胞与原代的差异。方法:用5个月人胎关节软骨分离培养细胞,观察细胞存活率、贴壁率、生长曲线和组织形态学的改变。结果:(1)软骨块在4℃下,3d 内细胞存活率可达 93.4％～97.6％。(2)原代细胞为圆形或三角形;第4代有一半转变成梭形,到第6代全部变为长梭形。(3)传代细胞贴壁时间(2～3h)短于原代(4～7h)。玻璃瓶内贴壁率传代细胞为78.7％～85.5％,原代8.8％。(4)生长曲线表明,从原代到第4代都有高增殖力;到第8代时增殖降低;到第12代几乎丧失细胞增殖。结论:软骨块4℃冷藏,3d内对细胞存活率无明显影响。第4代细胞贴壁快、增殖快,适于作为组织工程用细胞。第8代以后不适合。

Objective:To investigate the diffenrences between the primary chondrocytes and the sub-cultured ones. Methods: Five-month human fetal articular chondrocytes were isolated and cultured to ob-serve cell viability, cell adhesive rate, growth curves and morphological changes. Results: (1) When thelumps of cartilage were stored at 4℃, the viability of the chondrocytes was 97.6% to 93.4% within 3days; (2)The primary cells were spherical. But the subcultured cells gradually altered their own shape tofibroblast-like. At the 6th passage all cells altered their original spherical shape to the fibroblast-like; (3)The passage cells of 78.7% to 85.5% attached the glass flask, but the primary only 8.8%. (4) Thegrowth curves showed that the proliferation of the primary to the fourth passage was powerful, and to the8th passage, decreased apparently, and to the 12th lost their proliferative ability. Conclusion:The viabilityof chondrocytes may be not disturbed while the harvested cartilafe is stored at 4℃ within 3 days. On thebasis of the chondrocytic attachment and proliferation the earlier passage chondrocytes (around the 4thpassage)may be superior to the primary in the cartilage tissue engineering. But the latter passage ones(be-yond the 8th passage)are not suitable to the eartilage tissue engineering.