CAJ | 학술논문

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24 h SD大鼠,断颈处死后同时取材骨骺生长板La Croix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5～6 d可达到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.
목적 탐색불동조직래원적골후간세포체외공동배양적가행성.방법 선취건강신생24 h SD대서,단경처사후동시취재골후생장판La Croix배연골막조직화골후생장판정지구연골조직,체외분리공동배양,채용면역조직화학FGFR-3염색,대세포진행감정；채용면역조직화학번홍화“O”-량록염색화Ⅱ형효원단극륭항체염색,검측세포생물학특성.결과 래원우불동조직적세포피차상용、량호생장.도치현미경하관찰원대배양약5～6 d가체도80％세포융합,면역조직화학FG-FR-3염색표체양성,세포염색부위재포막화포장,증실체외공동배양적세포위골후간세포.세포정번홍화“O”-량록염색양성:세포정홍색,무록색.세포정Ⅱ형효원단극륭항체염색양성:격광공취초현미경하견세포정현록색형광,염색부위위우포질중,세포핵불착색.번홍화“O”-량록염색표체양성화Ⅱ형효원단극륭항체염색표체양성,설명공동배양적골후간세포가이정상지분비특정성기질.결론 불동조직래원적골후간세포가이재체외상용생장,병보지량호적생물학특성.
Objective To explore the feasibility of co-culture of precartilaginous 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.