人退变椎间盘软骨终板干细胞多向分化的研究

Identity of Chondrogenic Stem Cells in the Degenerated Human Cartilage Endplate

目的:为了分离和鉴定人退变椎间盘软骨终板干细胞。方法:收集因腰椎间盘退变性疾病行腰椎间盘摘除术并植骨融合的标本。在解剖显微镜下清理软骨终板组织,并消化软骨终板,提取软骨终板细胞。获得的软骨终板细胞经过琼脂糖三维筛选系统培养后,选取细胞克隆团并进行体外扩增,扩增后的细胞行流式细胞术检测干细胞标志物证实退变软骨终板中存在干细胞。结果:共聚焦免疫荧光提示退变椎间盘软骨终板组织中存在干细胞标志物STRO1、CD105、CD73、CD90阳性的细胞。经琼脂糖三维培养基筛选的CESCs在免疫表型上符合干细胞标准。结论:在人退变椎间盘的软骨终板中存在具有多向分化潜能的干细胞。

Objective： To isolate and investigate chondrogenic stem cells in the degenerated human cartilage endplate. Methods： The CEP used in this study was obtained from seven patients who underwent posterior discectomy and fusion for lumbar degenerative disease. Surgically explanted CEP was cleaned from any adherent extraneous tissue under a sterilized dissecting microscope. The nucleus pulposus, annulus fibrosus, and subchondral bone tissues around blocks of CEP were removed. The cells were subcultured in the agarose culture, and the cell clones were selected and expanded in vitro for the assays of stem cell markers to confirm the existence of stem cells in the cartilage endplate. Results： There are cells in the degenerated cartilage endplate which are positive for stem cell markers such as STRO-1, CD105, CD73, CD90 by the confocal scan microscope. By agarose culture separation, CESCs were proved to belong to stem cells in Immunophenotyping. Conclusion： All the results in this study indicated that the CESCs existed in the degenerated cartilage endplate.