脂肪间质干细胞修复犬关节软骨损伤的研究

Study on Repairing Canine Articular Cartilage Injury with Adipose Derived Mesenchymal Stem Cells

以手术方法建立2对共4处犬关节软骨组织损伤;使用无血清细胞培养液为对照,以标记有绿色荧光蛋白的犬脂肪间充质干细胞进行移植治疗试验。病理模型犬进行常规饲养15周后,对病理模型进行取样并系统检测。结果显示,试验组较对照组有更大面积和体积的新生关节软骨组织。新生软骨样组织呈绿色荧光蛋白(GFP)阳性。试验组与对照组的新生组织HE染色显示,试验组的细胞外基质染色较对照组更加均一,基质切面更加平整光滑。透明软骨的标志性基因COL2a1的免疫荧光染色显示,试验组可见较强的COL2a1阳性染色,而对照组仅有少量微弱的阳性染色。实时荧光定量PCR检测结果显示,试验组的COL2a1含量显著高于对照组(P<0.05)。试验结果表明,直接移植原代犬脂肪间充质干细胞到关节腔可有效修复犬软骨组织缺损,为干细胞治疗临床软骨组织损伤提供了科学依据。

Two pairs of articular cartilage tissue injuries were established by surgery;Using serum-free cell culture medium as control,canine adipose derived mesenchymal stem cells(ADMSCs)labeled with green fluorescent protein were transplanted.After 15 weeks of routine feeding,the pathological model was sampled for detection.The experimental group had larger area and volume of new cartilage tissue than the control group.The green fluorescent protein of the new cartilage like tissue was positive.HE staining of new tissues in the test group and the control group showed that the extracellular matrix staining in the test group was more uniform than that in the control group,and the matrix section was flatter.The immunofluorescence staining of type II collagen showed that the test group had strong positive staining of typeⅡcollagen,while the control group had only a small amount of weak positive staining.The results of real-time fluorescent quantitative PCR showed that the content of COL2a1 in the test group was significantly higher than that in the control group(P<0.05).This experiment clarified the feasibility of directly transplanting primary canine ADMSCs into the joint cavity to repair cartilage tissue defects,and provided clinical evidences for the future treatment of cartilage tissue injury by stem cells.