Department of Orthopedics, First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning, People's Republic of China.
PLoS One. 2012;7(8):e42804. doi: 10.1371/journal.pone.0042804. Epub 2012 Aug 3.
Numerous researches demonstrated the possibility of derivation of Schwann-like (SC-like) cells in vitro from bone marrow stromal cells (BMSCs). However, the concentration of the induce factors were different in those studies, especially for the critical factors forskolin (FSK) and β-heregulin (HRG). Here, we used a new and useful method to build an integrated microfluidic chip for rapid analyses of the optimal combination between the induce factors FSK and HRG. The microfluidic device was mainly composed of an upstream concentration gradient generator (CGG) and a downstream cell culture module. Rat BMSCs were cultured in the cell chambers for 11 days at the different concentrations of induce factors generated by CGG. The result of immunofluorescence staining on-chip showed that the group of 4.00 µM FSK and 250.00 ng/ml HRG presented an optimal effect to promote the derivation of SC-like cells. Moreover, the optimal SC-like cells obtained on-chip were further tested using DRG co-culture and ELISA to detect their functional performance. Our findings demonstrate that SC-like cells could be obtained with high efficiency and functional performance in the optimal inducers combination.
许多研究表明,骨髓基质细胞(BMSCs)在体外有可能分化为雪旺样(SC-like)细胞。然而,在这些研究中,诱导因子的浓度是不同的,特别是对于关键因子forskolin(FSK)和β-神经生长因子(HRG)。在这里,我们使用一种新的、有用的方法构建了一个集成的微流控芯片,用于快速分析诱导因子 FSK 和 HRG 之间的最佳组合。微流控装置主要由上游浓度梯度发生器(CGG)和下游细胞培养模块组成。将大鼠 BMSCs 培养在细胞室内,在 CGG 产生的不同浓度的诱导因子作用下培养 11 天。芯片上的免疫荧光染色结果表明,4.00 µM FSK 和 250.00 ng/ml HRG 组呈现出最佳的促进 SC-like 细胞分化的效果。此外,对芯片上获得的最佳 SC-like 细胞进行了进一步的 DRG 共培养和 ELISA 检测,以检测其功能性能。我们的研究结果表明,在最佳诱导剂组合下,可以高效、功能性地获得 SC-like 细胞。