Liu Xia, Zhou Guang-Dong, Liu Wei, Cao Yi-Lin
Plastic Surgery Hospital, Chinese Academy of Medical Science, Beiing 100144, China.
Zhonghua Zheng Xing Wai Ke Za Zhi. 2010 May;26(3):215-20.
To study the role of the soluble factors secreted by tissue engineered cartilage in promoting bone marrow stromal cells (BMSCs) chondrogenesis as an important aspect.
Porcine BMSCs, chondrocytes and dermal fibroblasts were respectively in vitro expanded and then seeded onto the polyglycolic acid/polylactic acid (PGA/PLA) scaffold. After 3 days, they were indirectly co-cultured by transwell. BMSCs-scaffold constructs were co-cultured with chondrocytes-scaffold constructs as experiment group (Exp), while co-cultured with fibroblasts-scaffold constructs as control group. BMSCs with the same cell number were seeded onto the scaffolds as another control group. There were 3 specimens in each group. All specimens were harvested after in vitro indirect co-culture for 8 weeks. Gross observation, histology, immunohistochemistry and RT-PCR were used to evaluate the results.
The BMSCs-scaffold constructs co-cultured with chondrocytes-scaffold shrunk gradually during in vitro culture, but formed the mature lacuna structures and metachromatic matrices, collagen II expression could be observed by immunohistochemistry and RT-PCR examination. In the control group, the constructs shrunk greatly during in vitro culture and showed mainly fibrous tissue.
The soluble factors secreted by chondrocytes can solely induce chondrogenic differentiation of BMSCs and thus promote the in vitro chondrogenesis of BMSCs.
作为重要的一方面，研究组织工程软骨分泌的可溶性因子在促进骨髓间充质干细胞（BMSCs）软骨形成中的作用。
将猪BMSCs、软骨细胞和真皮成纤维细胞分别进行体外扩增，然后接种到聚乙醇酸/聚乳酸（PGA/PLA）支架上。3天后，通过Transwell进行间接共培养。将BMSCs-支架构建体与软骨细胞-支架构建体共培养作为实验组（Exp），而与成纤维细胞-支架构建体共培养作为对照组。将相同细胞数的BMSCs接种到支架上作为另一个对照组。每组有3个标本。所有标本在体外间接共培养8周后收获。采用大体观察、组织学、免疫组织化学和RT-PCR评估结果。
与软骨细胞-支架共培养的BMSCs-支架构建体在体外培养过程中逐渐收缩，但形成了成熟的陷窝结构和异染性基质，免疫组织化学和RT-PCR检测可观察到Ⅱ型胶原表达。在对照组中，构建体在体外培养过程中大幅收缩，主要表现为纤维组织。
软骨细胞分泌的可溶性因子可单独诱导BMSCs向软骨分化，从而促进BMSCs的体外软骨形成。
