CAJ | 학술논문

背景：软骨细胞通过自分泌及旁分泌的作用可以为滑膜间充质干细胞向软骨细胞分化提供所需的生长因子及微环境，三维条件下更有利于细胞的黏附增殖与分化。 　　目的：观察滑膜间充质干细胞与软骨细胞混合培养于壳聚糖/Ⅰ型胶原复合支架材料中向成软骨细胞分化的能力。方法：取SD大鼠滑膜组织及软骨组织，用酶消化法获得滑膜间充质干细胞及软骨细胞分别进行培养。取第3代滑膜间充质干细胞及第2代软骨细胞，将二者以1∶2的比例混合培养负载于壳聚糖/Ⅰ型胶原复合支架材料21 d，进行激光共聚焦扫描及免疫组织化学检测。 　　结果与结论：培养72 h后，扫描电镜观察细胞黏附于支架材料表面，并可见细胞分泌大量基质成分。培养21 d后，激光共聚焦扫描可见细胞在支架表面分布均匀，逐层扫描后细胞逐渐减少。免疫组织化学检测可见基质能被Ⅱ型胶原染色，细胞染色呈现棕黄色。结果表明壳聚糖/Ⅰ型胶原复合支架材料提供三维生长空间，利用软骨细胞分泌生长因子及细胞间的相互作用可以诱导滑膜间充质干细胞向软骨细胞分化。
배경：연골세포통과자분비급방분비적작용가이위활막간충질간세포향연골세포분화제공소수적생장인자급미배경，삼유조건하경유리우세포적점부증식여분화。 　　목적：관찰활막간충질간세포여연골세포혼합배양우각취당/Ⅰ형효원복합지가재료중향성연골세포분화적능력。방법：취SD대서활막조직급연골조직，용매소화법획득활막간충질간세포급연골세포분별진행배양。취제3대활막간충질간세포급제2대연골세포，장이자이1∶2적비례혼합배양부재우각취당/Ⅰ형효원복합지가재료21 d，진행격광공취초소묘급면역조직화학검측。 　　결과여결론：배양72 h후，소묘전경관찰세포점부우지가재료표면，병가견세포분비대량기질성분。배양21 d후，격광공취초소묘가견세포재지가표면분포균균，축층소묘후세포축점감소。면역조직화학검측가견기질능피Ⅱ형효원염색，세포염색정현종황색。결과표명각취당/Ⅰ형효원복합지가재료제공삼유생장공간，이용연골세포분비생장인자급세포간적상호작용가이유도활막간충질간세포향연골세포분화。
BACKGROUND:Articular chondrocytes with the ability of autocrine and paracrine can provide the growth factors and microenvironment for synovial mesenchymal stem cels differentiating into the chondrocyte. The three-dimensional scaffold could provide space for cels adhesion, proliferation and differentiation. OBJECTIVE: To study the ability of chondrogenesis by co-culturing synovial mesenchymal stem cels and chondrocytes under the three-dimensional condition. METHODS:The synovial membrane and articular cartilage were harvested from rat knee joint. The synovial mesenchymal stem cels and chondrocytes were obtained through the method of enzyme digestion. The passage 3 synovial mesenchymal stem cels and passage 2 chondrocytes were co-cultured in the chitosan/I colagen composite scaffolds at the ratio of 1:2. Then, the cels/scaffold composite was harvested to be examined morphologicaly, histologicaly and immunohistochemicaly after being cultured 21 days. The confocal laser was also employed to detect the cels distribution in the scaffold. RESULTS AND CONCLUSION: After being cultured 72 hours, it could be observed from the cels/scaffold composite examined through the scanning electron microscope that the cels adhered on the surface of the scaffold and extracelular matrix surrounding the cels was seen on the scaffold. After being cultured 21 days, it could be found through the confocal laser scanning that the cels were wel-distributed on the scaffold, and cels decreased gradualy. Type II colagen was positive in the extracelular matrix immunohistochamicaly. It suggested from this study that the synovial mesenchymal stem cels could be co-cultured with chondrocytes in the chitosan/I colagen composite scaffolds and have the ability of chondrogenesis differentiation.