Department of Orthopedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, 600 Yishan Road, Shanghai, 200233, China.
Stem Cell Res Ther. 2022 Jul 30;13(1):386. doi: 10.1186/s13287-022-03094-6.
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.
种子细胞是细胞基软骨组织再生的关键因素。单独培养软骨细胞或间充质干细胞有几个局限性。近年来，共培养策略提供了潜在的解决方案。在这项研究中，直接共培养大鼠肋软骨细胞（CCs）和人 Wharton 氏胶间充质干细胞（hWJMSCs）被评估为再生关节软骨的候选物。
将大鼠 CCs 直接与 hWJMSCs 在微球模型中以不同比例（3:1、1:1、1:3）共培养 21 天。单核培养微球作为对照。进行 RT-qPCR、生化分析、组织学染色和评估，以分析每组的软骨分化情况。1:1 比例共培养微球组与单核培养对照组一起植入大鼠股骨沟的骨软骨缺损中 4、8、12 周。然后进行宏观和组织学评估。
与大鼠 CCs 微球组相比，3:1 和 1:1 比例组表现出相似的细胞外基质产生，但肥大倾向较小。免疫化学染色得到了一致的结果。RT-PCR 分析表明共培养的大鼠 CCs 促进了软骨形成，而抑制了肥大基因的表达。然而，hWJMSCs 在软骨形成方面仅略有改善，而在肥大表达方面无明显差异。体内实验表明，所有微球均能填充缺损，但共培养微球表现出较少的肥大、更好的周围软骨整合和适当的软骨下骨重塑。
大鼠 CCs 和 hWJMSCs 的共培养在体外和体内均表现出稳定的软骨形成表型和减少的肥大倾向。这些结果表明这种共培养组合是关节软骨再生的有前途的候选物。
