Functional classification of bone marrow dtromal cells (BMSCs) using DNA microarray, and development ofculture methods which can maintain differentiation potential of BMSCs

人骨髓基质细胞(HBMSCs)具有良好的分化能力和增殖能力，是骨组织工程应用的重要来源。然而，尽管骨髓间充质干细胞具有潜力，但它们在标准培养条件下经过多次倍增过程后失去了分化潜力。我们假设地塞米松可以增强骨髓间充质干细胞对分化试剂的反应性，并且地塞米松的持续治疗将建立具有更高分化潜能的骨髓间充质干细胞。我们的研究结果表明，地塞米松处理的细胞显示出比对照组更高的成骨能力和成骨能力。为了利用细胞和多孔支架开发高效的骨再生，将足够数量的细胞引入多孔支架是必不可少的。为了提高细胞接种效率，我们使用低压条件和Devi…改进了先前报道的细胞接种技术。更需要一种简单的装置来实现这种方法。程序如下。在第一步中，将多孔β-Tcp块放入玻璃室中，在第二步中，通过注射器将腔内的空气抽走以形成真空。第三步，将细胞悬浮液缓慢注入腔室，保持低压状态，并将块浸泡在悬浮液中。在最后一步，阀门被释放，从而使腔内的压力恢复正常。与自然穿透(对照组)相比，该方法的细胞种植效率提高了3倍，并且用该方法制备的种植体的成骨能力是对照组的3倍以上。我们还开发并测试了一种使用等离子体将细胞种植到多孔植入物中的系统，期望由纤维蛋白原形成的纤维蛋白网络将在几分钟内容纳引入的细胞，并促进骨形成。将BMSCs悬浮在血浆中，并将其引入多孔支架中，几分钟内就形成了纤维蛋白凝胶。然后植入物被移植。与悬浮于培养液中的BMSCs相比，使用等离子制备的种植体显示出更多的骨形成。较少

Human bone marrow stromal cells (hBMSCs) are an attractive source for bone tissue engineering applications because of their differentiation capability and their proliferation capability. However, despite the potential of BMSCs, they lose their differentiation potential after multiple doubling processes under standard culture conditions1. We hypothesized that dexamethasone could augment the responsiveness of BMSCs to differentiation reagents, and continuous dexamethasone treatment would establish BMSCs with higher differentiation potentials. The results of our studies indicated that the dexamethasone-treated cells showed higher osteogenic capability and bone formation capability than the control group.2. To develop efficient bone regeneration using cells and porous scaffolds, it is essential to introduce an adequate number of cells into porous scaffolds. To improve cell seeding efficiency, we modified the previously reported cell seeding techniques using low-pressure conditions and devi … More sed a simple device for this method. The procedures are as follows. In the first step, porous β-TCP blocks are placed into a glass chamber, and in the second step, the air in the chamber is removed by a syringe to create a vacuum. In the third step, the cell suspension is slowly infused into the chamber, maintaining the low-pressure condition, and the blocks are soaked in the suspension. In the final step, the valve is released so that the pressure in the chamber normalizes. Compared to spontaneous penetration (control), cell seeding efficiencies by this method were three fold higher, and the implants prepared by the method showed bone formation more than three folds of the control.3. We also developed and tested a cell seeding system into porous implants using plasma, expecting that the fibrin network formed from fibrinogen would hold introduced cells within minutes and promote bone formation. BMSCs were suspended in plasma and introduced into porous scaffolds, and fibrin gel was formed within minutes. Then the implants were transplanted. The implants prepared using plasma showed significantly more bone formation than that by the implants using BMSCs suspended in culture medium. Less