摘要
目的软骨微环境对软骨形成具有重要作用。本实验探讨软骨细胞与成纤维细胞共培养体外构建软骨的可行性。方法分别培养猪软骨细胞与人成纤维细胞,将两种细胞按3:7(软骨细胞:成纤维细胞)比例混匀,以5．0×107／ml终浓度接种于聚羟基乙酸支架(PGA,直径9mm,高2mm)作为共培养组,相同终浓度单纯软骨细胞和单纯成纤维细胞分别接种于相同支架作为阳性对照及阴性对照。每组各接种3例标本,每例接种细胞悬液200μl。全部标本均体外培养12周后取材,通过大体观察、组织学及免疫组化等检测对构建软骨进行评价。结果各组细胞均与材料支架粘附良好。体外培养12周后,阳性对照组(软骨细胞组)及共培养组均形成成熟的软骨样组织,组织学及免疫组化显示有成熟的软骨陷窝样结构及Ⅱ型胶原表达。软骨细胞组在体外培养过程中能基本保持复合物初始的大小和形状,形成的软骨在组织学上也教为均匀一致。共培养组在培养过程中稍有缩小,在复合物的周边区域形成了均质的软骨样组织,但在近中心区域存在一定量的纤维性组织。阴性对照组(单纯成纤维细胞组)在体外培养过程中逐渐皱缩变形,未形成软骨样组织。结论软骨微环境在体外软骨分化及软骨形成中具有重要作用,软骨细胞与成纤维细胞体外共培养能形成软骨样组织。
Objective Chondrogenic microenvironments play a vital role in chondrogenesis. This study explored the feasibility of chondrogenesis in vitro by co-culture of fibroblasts and chondrocytes. Methods Porcine auricular chondrocytes and human fibroblasts were isolated and expanded in vitro respectively and then were mixed at the ratio of 3:7 (chondrocytes: fibroblasts). The mixed cells were seeded onto polyglycolic acid (PGA)scaffold (9mm in diameter, 2mm in thickness)at the ultimate concentration of 5.0×107cells/ml as co-culture group. Chondrocytes and fibroblasts in the same ultimate concentration were seeded respectively onto the scaffolds as positive control group (chondrocytes group) and negative control group (fibroblasts group). The specimens were harvested after in vitro culture for 12weeks. Gross observation, histology and immunohistochemistry were used to evaluate the results. Results Cells in all groups had fine adhesion to the scaffold and could secrete abundant extracellular matrix. At 12 weeks, the specimens in both positive control group and co-culture group formed cartilage -like tissue with mature cartilage lacuna and typeⅡcollagen expression in histology and immunohistochemistry. In positive control group, the cell-scaffold constructs could maintain the original size and shape during in vitro culture and showed homogenous mature cartilage-like tissue. In co-culture group, the constructs shrunk slightly and formed homogenous cartilage-like tissue in the peripheral area with fibrous tissue in the central area. In negative control group, the constructs deformed and shrunk gradually without cartilage-like tissue formation. Conclusion Engineered cartilage-like tissue could be constructed by co-culture of fibroblasts and chondrocytes on a biodegradable scaffold.
出处
《组织工程与重建外科杂志》
2005年第4期217-219,共3页
Journal of Tissue Engineering and Reconstructive Surgery
基金
国家高技术发展计划项目(2002AA205021)上海市"重中之重"重点学科基金上海市青年科技"启明星"计划资助项目(05QNX1427)
