Development of Cell Transplantation System Using A Closed Culture for Heart Diseases

在使用细胞处理的再生医学中，细胞培养系统是最高障碍之一。大多数培养细胞治疗的临床试验都需要巨大的细胞处理中心，以确保生产产品的质量。我们的研究对封闭式细胞培养瓶进行了检验，并在第一年取得了良好的可行性和安全性。该培养体系的分化能力和增殖能力与开放式培养体系相同。最初，我们计划在第二年使用封闭式细胞培养体系进行外周动脉疾病的临床试验。但是，第二年开始对细胞加工的规定会更严格，我们不能进行临床试验。我们关注的是体内环境中长期培养细胞的安全性，这也是本研究要在体外环境中考察的问题之一。特别是，长期培养后的致癌作用在体外是被公认的。对于细胞疗法，致癌的风险是被仔细考虑的。在Beagle犬身上，进行了自体骨髓间充质细胞心脏移植。在移植前，细胞已经培养了5~10代。术后1~6个月，取心脏进行组织病理学分析。未发现癌和异型增生。没有发现异型增生和炎症。中期细胞培养不能诱导致癌变化。我们的结果表明，传代10次以下的细胞不会有明显的致癌作用。目前，一家公司生产了一种用于完全封闭细胞培养系统的机器人培养系统的试验版。机器人战略将减少这一过程中的成本和劳动力。

In regenerative medicine using cell processing, a cell culture system is one of the highest obstacles. Most clinical trials in cultivated cell therapy needed huge cell processing center, which ensure the quality of manufacturing products. Our study examined the closed cell culture bottle, and resulted in good feasibility and safety in the first year. The differentiation capability and proliferation ability in this culture system were the same as those cultivated in open culture systems.Initially, we planned the clinical trial for peripheral arterial disease using the close cell culture system in the second year. However, the regulation for the cell processing would be stricter in the start of the second year, and we could not perform the clinical trial. We have focused in the safety of long-term cultivated cells in vivo environment, which is one to be examined in vitro environment in this study. Especially, carcinogenesis following the long-term culture is recognized in vitro. For cell therapy, the risk of carcinogenesis is carefully considered. In beagle dogs, autologous bone marrow-derived mesenchymal cell transplantation to the heart was performed. The cells had been cultivated for 5 to 10 passages before grafting. After 1 to 6 months, the histopathological analyses of the hearts were carried out. Neither carcinoma nor dysplasia was recognized. Neither heteroplasia nor inflammation was recognized.Mid-term cell culture could not induce carcinogenic changes. Our results suggest that cells passaged under 10 times would be not significant carcinogenic. At present, a company produced a robotic culture system for completely closed cell culture system in a trial version. Robotics strategy would reduce the cost and labor in this process.