Shi Sirong, Xie Jing, Zhong Juan, Lin Shiyu, Zhang Tao, Sun Ke, Fu Na, Shao Xiaoru, Lin Yunfeng
State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, 610041, China.
Cell Prolif. 2016 Jun;49(3):341-51. doi: 10.1111/cpr.12259. Epub 2016 Apr 19.
Moving towards development of optimized cartilage regeneration with adipose-derived stromal cells (ASCs), the focus of this study was on investigating the influence of hypoxia on soluble factors secreted by ASCs and chondrocytes after crosstalk.
We established direct contact co-culture and non-contact co-culture systems by using red or green fluorescent protein (R/GFP)-labelled mice and SD rats respectively. Gene variation of growth factors of the two cell types, in both hypoxic and normoxic conditions, were screened using semi-quantitative polymerase chain reaction (PCR).
Co-culture with ASCs and chondrocytes under hypoxia was shown to successfully induce or enhance ASC to chondrogenic differentiation. To be specific, chondrogenic maker genes: AGC, COL II and SOX9 were remarkably enhanced in both ASCs and chondrocytes after crosstalk under low oxygen tension. Subsequently, screening growth factors in ASCs and chondrocytes under hypoxia showed that HIF-1α, VEGF-A/B, BMP-2/-4/-6, FGF-2 and IGF-1 were significantly increased, but not TGF-β1.
These results revealed that both hypoxia and co-culture systems can notably enhance chondrogenesis of ASCs as well as increase proliferation of ASCs and chondrocytes.
为了朝着利用脂肪来源的基质细胞（ASC）优化软骨再生的方向发展，本研究的重点是研究缺氧对ASC与软骨细胞相互作用后分泌的可溶性因子的影响。
我们分别使用红色或绿色荧光蛋白（R/GFP）标记的小鼠和SD大鼠建立了直接接触共培养和非接触共培养系统。在缺氧和常氧条件下，使用半定量聚合酶链反应（PCR）筛选两种细胞类型生长因子的基因变化。
缺氧条件下ASC与软骨细胞共培养显示成功诱导或增强了ASC向软骨细胞的分化。具体而言，在低氧张力下相互作用后，ASC和软骨细胞中的软骨生成标记基因：AGC、COL II和SOX9均显著增强。随后，对缺氧条件下ASC和软骨细胞中的生长因子进行筛选，结果显示HIF-1α、VEGF-A/B、BMP-2/-4/-6、FGF-2和IGF-1显著增加，但TGF-β1未增加。
这些结果表明，缺氧和共培养系统均可显著增强ASC的软骨生成能力，并增加ASC和软骨细胞的增殖。
