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目的 探讨晚期骨关节炎患者膝关节滑膜间质干细胞(synovium-derived mesenchymalstem cells,SMSCs)体外分离、培养的可行性及其在体外向脂肪细胞、成骨细胞和软骨细胞定向分化的特性.方法 取膝关节滑膜组织,胶原酶消化获得有核细胞.挑选单细胞克隆,筛选获得SMSCs.流式细胞技术检测细胞表面特异性抗原标志.培养至第三代,分别向脂肪细胞、成骨细胞和软骨细胞诱导分化.油红O染色鉴定向脂肪细胞分化;碱性磷酸酶染色、茜素红染色鉴定向成骨细胞分化;甲苯胺蓝染色鉴定向软骨细胞分化.RT-PCR检测脂肪细胞、成骨细胞标志基因.Ⅱ型胶原免疫组化染色检测软骨细胞Ⅱ型胶原的表达.结果 原代SMSCs体外培养呈葵花样细胞集落,传代后可见圆形巨噬样细胞和纺锤形成纤维样细胞,融合后呈成纤维细胞样生长.CD44、CD90呈阳性,CD34、CD71和CD45呈阴性.向脂肪细胞诱导21d,油红O染色阳性;RT-PCR检测有脂蛋白酶、乙二腈及PPARγ2表达;向成骨细胞诱导7、28 d,ALP,茜素红染色阳性,有ALP、Osteopontin及Osteocalcin表达;向软骨细胞诱导21d,甲苯胺蓝染色阳性,Ⅱ型胶原免疫组化染色阳性.结论 晚期骨关节炎患者膝关节滑膜组织可以分离、培养获得SMSCs. SMSCs具有向脂肪细胞、成骨细胞和软骨细胞发生定向分化的潜能.“,”Objective To study the method of isolating and culturing human synovium-derived mes-enchymal stem cels (SMSCs) from osteoarthritic knee joints and to investigate whether SMSCs could differ-entiate into osteoblasts, adipocytes and chondrocytes in vitro. Methods Cell populations were enzymatieally released from the synovial membrane obtained from human knee joints with end-stage osteoarthritis. Mono-clone was obtained by seleeting as primary SMSCs. SMSCs were characterized by FACS-analysis. SMSCs at passage 3 were used to investigate their multi-lineage potential in vitro. Upon treatments, phenotypes of cell cultures were analyzed by histo- and immunohistochemistry and by semi-quantitative reverse transcription-polymerase ehain reaction for the expression of lineage-related markers. Results Primary SMSCs, showed polygon and star shape. After passaged, it exhibited round eels and spindle cells. Consequently, homoge-neous populations of fibroblast-like cells were observed. SMSCs were positive for CD90 and CD44, negative for CD34, CD45 and CD71, respectively. Under appropriate euhure conditions, SMSCs were induced to dif-ferentiate to the adipocyte, osteocyte, and chondrocyte lineages, which were stained positively by Oil Red O staining, ALP staining, Alizarin red staining and Toluidine blue staining, respectively. Expression of adi-pogenic markers (Lipoprotein lipase, Adiponectin and PPARγ2) and osteogenic markers (ALP, Osteopontin and Osteocalein) were all detected. Type Ⅱ collagen was stained positively by immunohistochemistry stain-ing. Conclusion Human SMSCs can be isolated from knee joints of end-stage OA. These cells have the a-bility to proliferate extensively in culture, and they have the potential to differentiate into adipogenic, os-teogenic and chondrogenic lineage in vitro.