Automated mechanical passaging:: A novel and efficient method for human embryonic stem cell expansion
Automated mechanical passaging:: A novel and efficient method for human embryonic stem cell expansion
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DOI：
10.1634/stemcells.2005-0243
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发表时间：
2006-02-01
期刊：
影响因子：
5.2
通讯作者：
Chandran, Siddharthan
中科院分区：
文献类型：
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作者：
Joannides, Alexis;Fiore-Heriche, Christelle;Chandran, Siddharthan
人类胚胎干细胞（hESC）培养物的维持和繁殖需要更加标准化的方法。酶传代是目前应用最广泛的hESCs扩增方法。虽然快速和直接，这种技术导致大小不一的细胞团和显著的细胞创伤，这可能在长期培养中施加选择压力。机械传代具有确定菌落片段大小、减少细胞创伤和选择未分化菌落进行转移的可能性的潜在优势。然而，手工分离单个菌落是一个非常耗时的过程，不适合在不使用额外化学手段的情况下维持大量hESCs。在这项研究中，我们报告了一种高效的机械传代hESCs的自动化方法。我们专门使用这种方法在不使用酶消化的长期未分化培养中维持hESCs超过100天。因此，这种自动化技术可常规用于实验室培养hESCs。
There is a need for more standardized methods of maintenance and propagation of human embryonic stem cell (hESC) cultures. Enzymatic passaging currently represents the most widely used method for expansion of hESCs. Although rapid and straightforward, this technique results in variable-sized cell clusters and significant cellular trauma, which may apply selective pressure in long-term culture. Mechanical passaging has the potential advantages of defined colony fragment sizes, reduced cellular trauma, and the possibility of selecting undifferentiated colonies for transfer. However, manual dissection of individual colonies is a prohibitively time-consuming process unsuitable for maintaining large numbers of hESCs without the use of additional chemical means. In this study we report an efficient automated method for mechanically passaging hESCs. We have used this method exclusively to maintain hESCs in long-term undifferentiated culture without the use of enzymatic digestion for longer than 100 days. This automated technique can thus be used routinely to culture hESCs in the laboratory.