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

利用DNA微阵列对骨髓间充质干细胞（BMSCs）进行功能分类，并开发维持BMSCs分化潜能的培养方法

基本信息

批准号：
17390414
负责人：
SHINOMIYA Kenichi
金额：
$ 10.63万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2007
项目状态：
已结题

项目摘要

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

人骨髓基质细胞 (hBMSC) 因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，它们会失去分化潜力。培养地塞米松可以增强 BMSCs 对分化试剂的反应性，并且连续地塞米松治疗将建立具有更高分化潜力的 BMSCs。地塞米松处理的细胞比对照组表现出更高的成骨能力和骨形成能力。2.为了利用细胞和多孔支架开发有效的骨再生，必须将足够数量的细胞引入多孔支架中以提高细胞接种效率。我们使用低压条件修改了先前报道的细胞接种技术，并为该方法设计了一个简单的装置。第一步，将多孔 β-TCP 块放入玻璃室中。第二步，用注射器抽出腔室中的空气，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，将细胞块浸泡在悬浮液中。在最后一步中，释放阀门，使腔室中的压力恢复正常，与自发渗透（对照）相比，这种方法的细胞接种效率高出三倍，并且通过该方法制备的植入物显示出超过三倍的骨形成。的褶皱3.我们还开发并测试了使用血浆的细胞接种系统，期望由纤维蛋白原形成的纤维蛋白网络能够在几分钟内容纳引入的细胞并促进骨形成悬浮在血浆中并引入多孔支架中。纤维蛋白凝胶在几分钟内形成，然后移植使用血浆制备的植入物比使用悬浮在培养基中的骨髓间充质干细胞制备的植入物显示出明显更多的骨形成。