摘要
目的探索不同组织来源的骨骺干细胞体外共同培养的可行性。方法选取健康新生24hSD大鼠，断颈处死后同时取材骨骺生长板LaCroix环软骨膜组织和骨骺生长板静止区软骨组织，体外分离共同培养，采用免疫组织化学FGFR-3染色，对细胞进行鉴定；采用免疫组织化学蕃红花“O”-亮绿染色和Ⅱ型胶原单克隆抗体染色，检测细胞生物学特性。结果来源于不同组织的细胞彼此相容、良好生长。倒置显微镜下观察原代培养约5～6dN达到80％细胞融合，免疫组织化学FG—FR-3染色表达阳性，细胞染色部位在胞膜和胞浆，证实体外共同培养的细胞为骨骺干细胞。细胞呈蕃红花“O”-亮绿染色阳性：细胞呈红色，无绿色。细胞呈Ⅱ型胶原单克隆抗体染色阳性：激光共聚焦显微镜下见细胞呈现绿色荧光，染色部位位于胞质中，细胞核不着色。蕃红花“O”-亮绿染色表达阳性和Ⅱ型胶原单克隆抗体染色表达阳性，说明共同培养的骨骺干细胞可以正常地分泌特征性基质。结论不同组织来源的骨骺干细胞可以在体外相容生长，并保持良好的生物学特性。
Objective To explore the feasibility of co-culture of preeartilaginous stem cells （PSCs） coming from different tissues. Methods Cells isolated from La Croix and resting chondrocytes of growth plate of 24-hours old SD rats were co-cultured and identified by the expression of fibroblast growth factor receptor-3 （FGFR-3）. The biological characteristics of the cells were detected by immunocytochemistry Safranin＂O＂staining and collagen type II monoclonal antibody staining. Results The cells which came from La Croix and resting chondrocyte of growth plate were successfully differentiated and proliferated in vitro. The cells grew well and formed confluent growth on day 5-6. Positive expression of FGFR-3 located in the cell membrane and cytoplasm confirmed that the cocultured cells were precartilaginous stem cells （PSCs）. The cells were red, and not green with positive expression of Safranin＂O＂ staining. Positive expression of collagen type II monoclonal antibody was located in the cytoplasm with green fluorescence under laser scanning confocal microscope. It confirmed that co：cultured PSCs can secret characteristic matrix normally. Conclusions PSCs＇ which come from different tissues have good biocompatibility and can retain good biological characteristics.
出处
《中华小儿外科杂志》
CSCD
北大核心
2014年第1期66-69,共4页
Chinese Journal of Pediatric Surgery
基金
中国博士后科学基金第52批面上资助项目（2012M52121）5）
浙江省教育厅科研项目（Y201016893）
浙江中医药大学科研基金重点资助项目（2010ZZ02）
关键词
干细胞
骨骺
组织工程
Stem cells
Epiphyses
Tissue engineering
