Abstract：
Seeding cells are key factors in cell-based cartilage tissue regeneration. Monoculture of either chondrocyte or mesenchymal stem cells has several limitations. In recent years, co-culture strategies have provided potential solutions. In this study, directly co-cultured rat costal chondrocytes (CCs) and human Wharton\'s jelly mesenchymal stem (hWJMSCs) cells were evaluated as a candidate to regenerate articular cartilage.
Rat CCs are directly co-cultured with hWJMSCs in a pellet model at different ratios (3:1, 1:1, 1:3) for 21 days. The monoculture pellets were used as controls. RT-qPCR, biochemical assays, histological staining and evaluations were performed to analyze the chondrogenic differentiation of each group. The 1:1 ratio co-culture pellet group together with monoculture controls were implanted into the osteochondral defects made on the femoral grooves of the rats for 4, 8, 12 weeks. Then, macroscopic and histological evaluations were performed.
Compared to rat CCs pellet group, 3:1 and 1:1 ratio group demonstrated similar extracellular matrix production but less hypertrophy intendency. Immunochemistry staining found the consistent results. RT-PCR analysis indicated that chondrogenesis was promoted in co-cultured rat CCs, while expressions of hypertrophic genes were inhibited. However, hWJMSCs showed only slightly improved in chondrogenesis but not significantly different in hypertrophic expressions. In vivo experiments showed that all the pellets filled the defects but co-culture pellets demonstrated reduced hypertrophy, better surrounding cartilage integration and appropriate subchondral bone remodeling.
Co-culture of rat CCs and hWJMSCs demonstrated stable chondrogenic phenotype and decreased hypertrophic intendency in both vitro and vivo. These results suggest this co-culture combination as a promising candidate in articular cartilage regeneration.
Rat CCs are directly co-cultured with hWJMSCs in a pellet model at different ratios (3:1, 1:1, 1:3) for 21 days. The monoculture pellets were used as controls. RT-qPCR, biochemical assays, histological staining and evaluations were performed to analyze the chondrogenic differentiation of each group. The 1:1 ratio co-culture pellet group together with monoculture controls were implanted into the osteochondral defects made on the femoral grooves of the rats for 4, 8, 12 weeks. Then, macroscopic and histological evaluations were performed.
Compared to rat CCs pellet group, 3:1 and 1:1 ratio group demonstrated similar extracellular matrix production but less hypertrophy intendency. Immunochemistry staining found the consistent results. RT-PCR analysis indicated that chondrogenesis was promoted in co-cultured rat CCs, while expressions of hypertrophic genes were inhibited. However, hWJMSCs showed only slightly improved in chondrogenesis but not significantly different in hypertrophic expressions. In vivo experiments showed that all the pellets filled the defects but co-culture pellets demonstrated reduced hypertrophy, better surrounding cartilage integration and appropriate subchondral bone remodeling.
Co-culture of rat CCs and hWJMSCs demonstrated stable chondrogenic phenotype and decreased hypertrophic intendency in both vitro and vivo. These results suggest this co-culture combination as a promising candidate in articular cartilage regeneration.
摘要：
种子细胞是基于细胞的软骨组织再生的关键因素。软骨细胞或间充质干细胞的单一培养具有若干局限性。近年来,共同文化战略提供了潜在的解决方案。在这项研究中,直接共培养的大鼠肋软骨细胞（CC）和人沃顿胶质间充质干细胞（hWJMSCs）被评估为再生关节软骨的候选细胞。
将大鼠CC与hWJMSC在颗粒模型中以不同比例(3:1、1:1、1:3)直接共培养21天。将单一培养物颗粒用作对照。RT-qPCR,生化化验,进行组织学染色和评估以分析各组的软骨形成分化。将1：1比例的共培养颗粒组与单培养对照一起植入在大鼠股骨沟上形成的骨软骨缺损中4、8、12周。然后，进行宏观和组织学评估.
与大鼠CCs颗粒组相比，3：1和1：1比率组显示出相似的细胞外基质产生，但肥大倾向较低。免疫化学染色发现一致的结果。RT-PCR分析表明，共培养的大鼠CC促进了软骨形成，而肥大基因的表达受到抑制。然而,hWJMSCs在软骨形成方面仅显示出轻微改善，但在肥大表达方面没有显着差异。体内实验表明，所有的颗粒填充的缺陷，但共培养颗粒显示减少肥大，更好的周围软骨整合和适当的软骨下骨重塑。
大鼠CC和hWJMSCs的共培养在体外和体内都显示出稳定的软骨形成表型和降低的肥大强度。这些结果表明这种共培养组合是关节软骨再生中的有希望的候选物。
将大鼠CC与hWJMSC在颗粒模型中以不同比例(3:1、1:1、1:3)直接共培养21天。将单一培养物颗粒用作对照。RT-qPCR,生化化验,进行组织学染色和评估以分析各组的软骨形成分化。将1：1比例的共培养颗粒组与单培养对照一起植入在大鼠股骨沟上形成的骨软骨缺损中4、8、12周。然后，进行宏观和组织学评估.
与大鼠CCs颗粒组相比，3：1和1：1比率组显示出相似的细胞外基质产生，但肥大倾向较低。免疫化学染色发现一致的结果。RT-PCR分析表明，共培养的大鼠CC促进了软骨形成，而肥大基因的表达受到抑制。然而,hWJMSCs在软骨形成方面仅显示出轻微改善，但在肥大表达方面没有显着差异。体内实验表明，所有的颗粒填充的缺陷，但共培养颗粒显示减少肥大，更好的周围软骨整合和适当的软骨下骨重塑。
大鼠CC和hWJMSCs的共培养在体外和体内都显示出稳定的软骨形成表型和降低的肥大强度。这些结果表明这种共培养组合是关节软骨再生中的有希望的候选物。
