中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
1期
100-105
,共6页
邵博%龚忠诚%刘慧%凌彬%克热木%尹小朋%胡露露%王冰%宁晓婷%林兆全
邵博%龔忠誠%劉慧%凌彬%剋熱木%尹小朋%鬍露露%王冰%寧曉婷%林兆全
소박%공충성%류혜%릉빈%극열목%윤소붕%호로로%왕빙%저효정%림조전
干细胞%诱导%滑膜间充质干细胞%软骨形成%软骨细胞%可溶性因子%生长因子%种子细胞%三维培养%新疆维吾尔自治区自然科学基金
榦細胞%誘導%滑膜間充質榦細胞%軟骨形成%軟骨細胞%可溶性因子%生長因子%種子細胞%三維培養%新疆維吾爾自治區自然科學基金
간세포%유도%활막간충질간세포%연골형성%연골세포%가용성인자%생장인자%충자세포%삼유배양%신강유오이자치구자연과학기금
chondrocytes%synovial membrane%mesenchymal stem cells%cellculture%celldifferentiation%tissue engineering
背景：滑膜间充质干细胞在体外具有多向分化的能力，有望成为软骨组织工程中治疗软骨缺损的种子细胞，在其向软骨细胞分化过程中，合适的生长因子起了重要作用。<br> 目的：利用富含生长因子的软骨细胞上清液诱导滑膜间充质干细胞向软骨细胞分化，并对其鉴定。<br> 方法：采用消化法分别获得 SD 大鼠滑膜间充质干细胞、软骨细胞。收集软骨细胞上清液离心、过滤冻存备用。培养滑膜间充质干细胞至第3代后离心成微团，并用软骨细胞上清液进行成软骨诱导分化，通过形态学观察、免疫组织化学法、RT-PCR检测进行鉴定。<br> 结果与结论：滑膜间充质干细胞使用软骨细胞上清液成软骨诱导21 d后，微团可见似软骨样组织。免疫组化法进行Ⅱ型胶原鉴定，基质能被Ⅱ型胶原染色，细胞染色呈现棕黄色。RT-PCR 结果显示诱导后的微团表达软骨特异性基因Ⅱ型胶原和蛋白聚糖。证实软骨细胞分泌的可溶性因子可以诱导大鼠滑膜间充质干细胞向软骨方向分化。
揹景：滑膜間充質榦細胞在體外具有多嚮分化的能力，有望成為軟骨組織工程中治療軟骨缺損的種子細胞，在其嚮軟骨細胞分化過程中，閤適的生長因子起瞭重要作用。<br> 目的：利用富含生長因子的軟骨細胞上清液誘導滑膜間充質榦細胞嚮軟骨細胞分化，併對其鑒定。<br> 方法：採用消化法分彆穫得 SD 大鼠滑膜間充質榦細胞、軟骨細胞。收集軟骨細胞上清液離心、過濾凍存備用。培養滑膜間充質榦細胞至第3代後離心成微糰，併用軟骨細胞上清液進行成軟骨誘導分化，通過形態學觀察、免疫組織化學法、RT-PCR檢測進行鑒定。<br> 結果與結論：滑膜間充質榦細胞使用軟骨細胞上清液成軟骨誘導21 d後，微糰可見似軟骨樣組織。免疫組化法進行Ⅱ型膠原鑒定，基質能被Ⅱ型膠原染色，細胞染色呈現棕黃色。RT-PCR 結果顯示誘導後的微糰錶達軟骨特異性基因Ⅱ型膠原和蛋白聚糖。證實軟骨細胞分泌的可溶性因子可以誘導大鼠滑膜間充質榦細胞嚮軟骨方嚮分化。
배경：활막간충질간세포재체외구유다향분화적능력，유망성위연골조직공정중치료연골결손적충자세포，재기향연골세포분화과정중，합괄적생장인자기료중요작용。<br> 목적：이용부함생장인자적연골세포상청액유도활막간충질간세포향연골세포분화，병대기감정。<br> 방법：채용소화법분별획득 SD 대서활막간충질간세포、연골세포。수집연골세포상청액리심、과려동존비용。배양활막간충질간세포지제3대후리심성미단，병용연골세포상청액진행성연골유도분화，통과형태학관찰、면역조직화학법、RT-PCR검측진행감정。<br> 결과여결론：활막간충질간세포사용연골세포상청액성연골유도21 d후，미단가견사연골양조직。면역조화법진행Ⅱ형효원감정，기질능피Ⅱ형효원염색，세포염색정현종황색。RT-PCR 결과현시유도후적미단표체연골특이성기인Ⅱ형효원화단백취당。증실연골세포분비적가용성인자가이유도대서활막간충질간세포향연골방향분화。
BACKGROUND:Synovial mesenchymal stem cells have the ability of multilineage differentiation in vitro, which are expected to be seed cells for the treatment of cartilage defects in cartilage tissue engineering. Appropriate growth factors are critical for the chondrocyte differentiation of synovial mesenchymal stem cells. <br> OBJECTIVE:To study the role of secreted factors by chondrocytes to induce chondrogenesis of synovial mesenchymal stem cells. <br> METHODS:The synovial mesenchymal stem cells and chondrocytes were harvested from rat knee joints and cultured through the digestion method. The supernatant was col ected from chondrocytes, and centrifuged, filtered and cryopreserved. The third passage synovial mesenchymal stem cells centrifuged as pel ets were cultured in the chondrocyte supernatant for 21 days. And the cells morphology was examined and the type II col agen and aggrecan were detected through immunohistochemistry and RT-PCR. <br> RESULTS AND CONCLUSION:The synovial mesenchymal stem cellpel ets cultured in the chondrocyte supernatant became cartilage-like tissue after 21 days. The type II col agen was detected positively in the matrix of synovial mesenchymal stem cellpel et immunohistochemical y. RT-PCR examination showed that the type II col agen and aggrecan expressed in the synovial mesenchymal stem cellpel et cultured in the chondrocyte supernatant. It suggested that synovial mesenchymal stem cellcould be induced to differentiate into chondrocytes depending on soluble factors secreted by chondrocytes.
