State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University , Chengdu 610041, China.
Institute of Stomatology, General Hospital of Chinese PLA , Beijing 100853, China.
Bone Res. 2016 Feb 3;4:15036. doi: 10.1038/boneres.2015.36. eCollection 2016.
Adipose-derived stem cells (ASCs) and mesenchymal stem cells are promising for tissue repair because of their multilineage differentiation capacity. Our previous data confirmed that the implantation of mixed ASCs and chondrocytes into cartilage defects induced desirable in vivo healing outcomes. However, the paracrine action of ASCs on chondrocytes needs to be further elucidated. In this study, we established a co-culture system to achieve cell-to-cell and cell-to-tissue crosstalk and explored the soluble growth factors in both ASCs and chondrocytes supplemented with 1% fetal bovine serum to mimic the physiological microenvironment. In ASCs, we screened for growth factors by semi-quantitative PCR and quantitative real-time PCR and found that the expression of bone morphogenetic protein 2 (BMP-2), vascular endothelial growth factor B (VEGFB), hypoxia inducible factor-1α (HIF-1α), fibroblast growth factor-2 (FGF-2), and transforming growth factor-β1 significantly increased after co-culture in comparison with mono-culture. In chondrocytes, VEGFA was significantly enhanced after co-culture. Unexpectedly, the expression of collagen II and aggrecan was significantly down-regulated in the co-culture group compared with the mono-culture group. Meanwhile, among all the growth factors screened, we found that the BMP family members BMP-2, BMP-4, and BMP-5 were down-regulated and that VEGFB, HIF-1α, FGF-2, and PDGF were significantly decreased after co-culture. These results suggest that crosstalk between ASCs and chondrocytes is a pathway through the regulated growth factors that might have potential in cartilage repair and regeneration and could be useful for tissue engineering.
脂肪来源干细胞(ASCs)和间充质干细胞因其多能分化能力而在组织修复中具有广阔的应用前景。我们之前的数据证实,将混合 ASC 和软骨细胞植入软骨缺损中,可诱导出理想的体内愈合效果。然而,ASCs 对软骨细胞的旁分泌作用仍需要进一步阐明。在本研究中,我们建立了共培养体系,以实现细胞间和细胞与组织的串扰,并探讨了在补充 1%胎牛血清的情况下 ASC 和软骨细胞中的可溶性生长因子,以模拟生理微环境。在 ASC 中,我们通过半定量 PCR 和定量实时 PCR 筛选生长因子,发现与单核培养相比,共培养后骨形态发生蛋白 2(BMP-2)、血管内皮生长因子 B(VEGFB)、缺氧诱导因子-1α(HIF-1α)、成纤维细胞生长因子 2(FGF-2)和转化生长因子-β1 的表达显著增加。在软骨细胞中,VEGFA 的表达在共培养后显著增强。出乎意料的是,与单核培养组相比,共培养组的 II 型胶原和聚集蛋白聚糖的表达显著下调。同时,在筛选出的所有生长因子中,我们发现 BMP 家族成员 BMP-2、BMP-4 和 BMP-5 的表达下调,而 VEGFB、HIF-1α、FGF-2 和 PDGF 的表达在共培养后显著降低。这些结果表明,ASCs 和软骨细胞之间的串扰是一种通过调节生长因子来实现的途径,可能在软骨修复和再生中具有潜力,并可能对组织工程有用。