Dahlin Rebecca L, Meretoja Ville V, Ni Mengwei, Kasper F Kurtis, Mikos Antonios G
Department of Bioengineering, Rice University , Houston, Texas.
Tissue Eng Part A. 2014 Nov;20(21-22):2883-91. doi: 10.1089/ten.TEA.2014.0107. Epub 2014 Jun 3.
This work investigated the effect of flow perfusion bioreactor culture with and without transforming growth factor-β3 (TGF-β3) supplementation on the proliferation, extracellular matrix (ECM) production, and chondrogenic gene expression of chondrocytes both in monoculture and in co-culture with bone marrow-derived mesenchymal stem cells (MSCs). Both cell populations were cultured on electrospun poly(ɛ-caprolactone) scaffolds for 2 weeks in static or flow perfusion culture with and without TGF-β3. Overall, it was observed that without growth factors, flow perfusion culture resulted in increased cell proliferation and ECM with a more cartilage-like composition. While with TGF-β3 induction, flow perfusion constructs generally had lower chondrogenic gene expression than the corresponding static cultures, the growth factor still had an inductive effect on the cells with enhanced gene expression compared with the corresponding noninduced cultures. In addition, while flow perfusion cultures generally had reduced overall ECM content, the ECM distribution was more homogenous compared with the corresponding static cultures. These results are significant in that they indicate that while flow perfusion culture has some beneficial effects on the chondrogenic phenotype of articular chondrocytes, flow perfusion alone is not sufficient to maintain the chondrogenic phenotype of chondrocytes in either monoculture or co-culture, thus demonstrating the advantages of using exogenously added growth factors in flow perfusion culture. Furthermore, the results demonstrate the advantages of flow perfusion culture for the creation of large tissue engineered constructs and the potential of co-cultures of articular chondrocytes and MSCs to be used in flow perfusion culture.
本研究探讨了在添加和不添加转化生长因子-β3（TGF-β3）的情况下，流动灌注生物反应器培养对软骨细胞在单培养以及与骨髓间充质干细胞（MSC）共培养时的增殖、细胞外基质（ECM）产生和软骨生成基因表达的影响。两种细胞群体均在静电纺丝聚（ε-己内酯）支架上培养2周，分别处于添加和不添加TGF-β3的静态或流动灌注培养条件下。总体而言，观察到在无生长因子的情况下，流动灌注培养导致细胞增殖增加以及ECM具有更类似软骨的组成。虽然在TGF-β3诱导下，流动灌注构建体的软骨生成基因表达通常低于相应的静态培养物，但与相应的未诱导培养物相比，生长因子对细胞仍具有诱导作用，可增强基因表达。此外，虽然流动灌注培养的总体ECM含量通常降低，但与相应的静态培养物相比，ECM分布更均匀。这些结果具有重要意义，因为它们表明，尽管流动灌注培养对关节软骨细胞的软骨生成表型有一些有益影响，但仅流动灌注不足以在单培养或共培养中维持软骨细胞的软骨生成表型，从而证明了在流动灌注培养中使用外源性添加生长因子的优势。此外，结果证明了流动灌注培养在创建大型组织工程构建体方面的优势，以及关节软骨细胞与MSC共培养在流动灌注培养中的应用潜力。
