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

人骨髓基质干细胞（hBMSCs）由于其分化能力和增殖能力而成为骨组织工程应用的一个有吸引力的来源。然而，尽管BMSC具有潜力，但在标准培养条件下经过多次倍增过程后，它们失去了分化潜力1。我们假设地塞米松可以增强BMSCs对分化试剂的反应性，并且连续的地塞米松处理将建立具有更高分化潜能的BMSCs。实验结果表明，地塞米松处理组的成骨能力和成骨能力均高于对照组.为了利用细胞和多孔支架开发有效的骨再生，将足够数量的细胞引入多孔支架是必要的。为了提高细胞接种效率，我们修改了以前报道的细胞接种技术，使用低压条件和设备， ...更多信息 为这种方法提供了一个简单的装置。程序如下。在第一步中，将多孔β-TCP块放入玻璃腔室中，在第二步中，通过注射器去除腔室中的空气以产生真空。在第三步中，将细胞悬浮液缓慢注入腔室中，保持低压条件，并将块浸泡在悬浮液中。在最后一步中，阀门被释放，使得腔室中的压力正常化。与自发穿透（对照）相比，通过该方法的细胞接种效率高三倍，并且通过该方法制备的植入物显示出超过对照三倍的骨形成.我们还开发并测试了使用血浆将细胞接种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保持引入的细胞并促进骨形成。骨髓基质细胞悬浮在血浆中，并引入到多孔支架，和纤维蛋白凝胶在几分钟内形成。然后移植植入物。使用血浆制备的植入物显示出比使用悬浮在培养基中的BMSC的植入物显着更多的骨形成。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