单层培养的人关节软骨细胞生物学行为研究

Study on biological behavior of human articular chondrocytes in monolayer culture

目的:研究单层培养的人关节软骨细胞生物学行为的改变。方法:对来自8例手术切除负重关节非负重区的软骨细胞在10%DMEM中进行单层传代培养,观察细胞形态学、增殖细胞倍增时间(DT)、细胞Ⅱ型胶原免疫组织化学染色以及流式细胞分析细胞增殖指数(PI)和细胞凋亡。结果:随着传代次数的增加,(1)细胞形态由原代的多角形转变为第3、4代的短梭形、第8、9代的成纤维细胞形态;(2)DT在逐渐延长,PI在逐渐减小;(3)细胞分泌的Ⅱ型胶原逐渐减少;(4)细胞凋亡指数未见明显的改变(P>0.05)。结论:体外单层培养的关节软骨细胞存在着失分化和老化,但第3、4代能较好地保持软骨细胞的生物学行为,可作为种子细胞的来源。

Objective： To study the changes of biological behavior of articular chondrocytes in monolayer culture. Methods： The chondrocytes from 8 human weight - loading articulations were cultured in monolayer and passaged in 10% DMEM, to observe their morphology, doubling time （DT）, immunohistochemical stain of collagen Ⅱ, proliferation index and apoptosis index by flow cytometer. Results：With the cells passaged, （1）their original polygonal shape changed to fusiform at 3rd and 4th passage and spindle at 8th or 9th passage; （ 2 ） DT was prolonging, PI decreasing gradually; （ 3 ） Collagen Ⅱ secretion decreasing; （4） apoptosis index changed little（ P 〉 0.05）. Conclusion： The chondrocytes saltured in monolayer may become dedifferentiation and aging, but the 3rd and 4th passage cells keep their fine biological behavior, may be served as seeding cells.