Wang Jingkai, Tao Yiqing, Zhou Xiaopeng, Li Hao, Liang Chengzhen, Li Fangcai, Chen Qi-Xin
Department of Orthopedics, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China.
Exp Biol Med (Maywood). 2016 Dec;241(18):2104-2111. doi: 10.1177/1535370216662362. Epub 2016 Aug 19.
Recent studies indicated that cell-based therapy could be a promising approach to treat intervertebral disc degeneration. Though the harsh microenvironment in disc is still challenging to implanted cells, it could be overcome by pre-conditioning graft cells before transplantation, suggested by previous literatures. Therefore, we designed this study to identify the potential effect of chondrogenic pre-differentiation on adipose-derived mesenchymal stem cells in intervertebral disc-like microenvironment, characterized by limited nutrition, acidic, and high osmosis in vitro. Adipose-derived mesenchymal stem cells of rat were divided into five groups, embedded in type II collagen scaffold, and cultured in chondrogenic differentiation medium for 0, 3, 7, 10, and 14 days. Then, the adipose-derived mesenchymal stem cells were implanted and cultured in intervertebral disc-like condition. The proliferation and differentiation of adipose-derived mesenchymal stem cells were evaluated by cell counting kit-8 test, real-time quantitative polymerase chain reaction, and Western blotting and immunofluorescence analysis. Analyzed by the first week in intervertebral disc-like condition, the results showed relatively greater proliferative capability and extracellular matrix synthesis ability of the adipose-derived mesenchymal stem cells pre-differentiated for 7 and 10 days than the control. We concluded that pre-differentiation of rat adipose-derived mesenchymal stem cells in chondrogenic culture medium for 7 to 10 days could promote the regeneration effect of adipose-derived mesenchymal stem cells in intervertebral disc-like condition, and the pre-differentiated cells could be a promising cell source for disc regeneration medicine.
近期研究表明，基于细胞的疗法可能是治疗椎间盘退变的一种有前景的方法。尽管椎间盘内恶劣的微环境对植入细胞来说仍然是一项挑战，但先前的文献表明，这一问题可通过在移植前对移植细胞进行预处理来克服。因此，我们设计了本研究，以确定在体外以营养有限、酸性和高渗透压为特征的椎间盘样微环境中，软骨形成预分化对脂肪间充质干细胞的潜在影响。将大鼠脂肪间充质干细胞分为五组，包埋于Ⅱ型胶原支架中，并在软骨分化培养基中培养0、3、7、10和14天。然后，将脂肪间充质干细胞植入并在椎间盘样条件下培养。通过细胞计数试剂盒-8检测、实时定量聚合酶链反应、蛋白质免疫印迹法和免疫荧光分析评估脂肪间充质干细胞的增殖和分化。在椎间盘样条件下培养第一周进行分析，结果显示，预分化7天和10天的脂肪间充质干细胞的增殖能力和细胞外基质合成能力相对高于对照组。我们得出结论，大鼠脂肪间充质干细胞在软骨分化培养基中预分化7至10天可促进脂肪间充质干细胞在椎间盘样条件下的再生效果，且预分化细胞可能是椎间盘再生医学中一种有前景的细胞来源。
