A Compact, Automated Cell Culture System for Clinical Scale Cell Expansion from Primary Tissues
A Compact, Automated Cell Culture System for Clinical Scale Cell Expansion from Primary Tissues
复制标题
DOI：
10.1089/ten.tec.2009.0305
复制
发表时间：
2010-10-01
影响因子：
3
通讯作者：
Kagami, Hideaki
中科院分区：
文献类型：
--
作者：
Kato, Ryuji;Iejima, Daisuke;Kagami, Hideaki
尽管临床实用的自动化细胞培养系统的数量不断增加，但对具有更大能力的更紧凑平台的需求也在增加，以直接从患者组织制备原代细胞。在这里，我们报告了一个自动化的细胞培养系统，也是紧凑的发展。该机器由一个供应单元、一个孵化单元和一个收集单元组成，这些单元可以容纳在70 cm x 60 cm x 86 cm的空间内。紧凑的尺寸是通过我们的概念实现的，即从原代培养步骤到最终细胞收获使用单个培养容器，而不是用多个培养容器按比例放大。人成纤维细胞和骨髓基质细胞（BMSCs）成功地培养与此系统超过19天无污染。从三片牙龈组织（2 mm x 2 mm）或从10 mL骨髓穿刺液中，该系统可以分别产生超过2.0 x 10（7）个细胞和高达3.0 x 10（7）个细胞的成纤维细胞和BMSC。该系统培养的骨髓间充质干细胞移植到裸鼠皮下后可异位成骨。我们的原型系统将为最大限度地减少具有临床相关细胞产量的自动培养机械提供基础，同时还将自动化能力扩展到包括原代组织培养。
Despite the growing number of clinically practical automated cell culture systems, demand is also increasing for more compact platforms with greater capabilities to prepare primary cells directly from patient tissue. Here we report the development of an automated cell culture system that is also compact. The machinery consisted of a supply unit, an incubation unit, and a collection unit, which fit within a 70 cm x 60 cm x 86 cm space. The compact size was enabled by our concept of using a single culture vessel from the primary culture steps to final cell harvest instead of scaling up with multiple culture vessels. Human fibroblasts and bone marrow stromal cells (BMSCs) were successfully cultured with this system over 19 days without contamination. From three pieces of gingival tissue (2 mm x 2 mm) or from 10 mL of bone marrow aspirate, the system could produce more than 2.0 x 10(7) cells and up to 3.0 x 10(7) cells for fibroblasts and BMSCs, respectively. The BMSCs produced by this system were capable of ectopic bone formation after transplantation into the subcutaneous space of nude mice. Our prototype system will provide a foundation for minimizing automatic culture machinery with clinically relevant cell yields while also expanding the automation capabilities to include primary tissue culture.