CAJ | 학술논문

背景：关节软骨损伤后修复结果不满意，需要新的手段，而脂肪间充质干细胞较适宜做种子细胞诱导软骨，然而怎么能够使诱导的软骨具有功能需要研究。目的：采用三维培养体系诱导人脂肪间充质干细胞微球向软骨分化。方法：无菌切取吸脂术后脂肪组织，分离培养人脂肪间充质干细胞，传至第3代进行流式细胞术分析，成骨成脂肪诱导等鉴定，同时也给予合适的培养条件用三维培养的方式向软骨细胞诱导，并行阿利辛蓝染色鉴定糖胺多糖的合成，苏木精-伊红染色进行组织学分析，免疫荧光检测Ⅱ型胶原表达，称质量测量软骨硬度。结果与结论：分离的人脂肪间充质干细胞CD105，CD44，CD29均高表达，而 CD45，CD34低表达，并且成骨成脂诱导后细胞茜素红染色和油红O染色均为阳性。三维培养法诱导的软骨细胞可表达大量糖胺多糖及Ⅱ型胶原。结果证实，三维培养法诱导人脂肪间充质细胞向软骨分化后，具有软骨细胞的特性。
배경：관절연골손상후수복결과불만의，수요신적수단，이지방간충질간세포교괄의주충자세포유도연골，연이즘요능구사유도적연골구유공능수요연구。목적：채용삼유배양체계유도인지방간충질간세포미구향연골분화。방법：무균절취흡지술후지방조직，분리배양인지방간충질간세포，전지제3대진행류식세포술분석，성골성지방유도등감정，동시야급여합괄적배양조건용삼유배양적방식향연골세포유도，병행아리신람염색감정당알다당적합성，소목정-이홍염색진행조직학분석，면역형광검측Ⅱ형효원표체，칭질량측량연골경도。결과여결론：분리적인지방간충질간세포CD105，CD44，CD29균고표체，이 CD45，CD34저표체，병차성골성지유도후세포천소홍염색화유홍O염색균위양성。삼유배양법유도적연골세포가표체대량당알다당급Ⅱ형효원。결과증실，삼유배양법유도인지방간충질세포향연골분화후，구유연골세포적특성。
BACKGROUND:Adipose-derived mesenchymal stem cels are more appropriate as seed cels for cartilage induction in patients who have unsatisfied repair after articular cartilage injuries, but how to induce the cartilage function need to be studied. OBJECTIVE:To induce the chondrogenic differentiation of human adipose-derived mesenchymal stem cel microspheres using three-dimensional culture system. METHODS:Adipose tissues were extracted after liposuction under sterile, to isolate and culture human adipose-derived mesenchymal stem cels. The third passage cels were analyzed by flow cytometry and identified by osteogenic induction and adipogenic induction, and meanwhile, cels were cultured under three-dimensional culture system for chondrogenic differentiation. Then, cels were observed by Alcian blue staining for identification of glycosaminoglycan synthesis as wel as hematoxylin-eosin staining for histological analysis. Expression of colagen type II was detected by immunofluorescence method. Cartilage stiffness was determined by mass measurement. RESULTS AND CONCLUSION:Isolated human adipose-derived mesenchymal stem cels highly expressed CD105, CD44, CD29, but showed low expressions of CD45, CD34. After osteogenic and adipogenic induction, cels were positive for alizarin red staining and oil red O staining. Under the three-dimensional culture, chondrocytes could greatly express glycosaminoglycans and colagen type II. The results suggest that under the three-dimensional culture, human adipose-derived mesenchymal cels differentiate into chondrocytes and have the characteristics of chondrocytes.