Synthetic Gene Regulation Circuits for Control of Cell Expansion
Synthetic Gene Regulation Circuits for Control of Cell Expansion
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DOI：
10.1089/ten.tea.2009.0184
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发表时间：
2010-02-01
影响因子：
4.1
通讯作者：
Wirth, Dagmar
中科院分区：
文献类型：
--
作者：
May, Tobias;Butueva, Milada;Wirth, Dagmar
在原代细胞分析和再生科学中的主要缺点涉及细胞的有限数量和同质性。这种限制可以通过保留感兴趣的细胞类型的特性的体外细胞扩增来克服。然而，对于大多数原代分化的细胞，增殖能力是有限的和/或增殖与细胞的去分化相关。我们开发了一种灵活的细胞扩增策略，可以严格可靠地控制细胞增殖。该系统依赖于采用基于四环素控制的正反馈回路的合成基因模块。构建这些基因模块并通过慢病毒载体转导。我们成功地产生了鼠内皮细胞系，重要的是也产生了人内皮细胞系。建立的细胞系的关键特征是它们的增殖状态可以被严格控制，同时维持相关标志物的表达。在细胞克隆和细胞库中观察到这种严格的增殖控制，甚至在同时使用两个独立的永生化基因时也能维持这种控制。因此，这种策略是灵活的，易于处理，可靠的。最重要的是，它允许扩增具有原代样表型的人细胞。
A major drawback in the analysis of primary cells and in regenerative sciences concerns the limited number and homogeneity of cells. This limitation could be overcome by in vitro cell expansion that retains the properties of the cell types of interest. However, for most primary differentiated cells the proliferation capacity is finite and/or proliferation is associated with dedifferentiation of cells. We have developed a flexible cell expansion strategy that allows strict and reliable control of cell proliferation. This system relies on synthetic gene modules that employ positive feedback loops based on Tetracycline control. These gene modules were constructed and transduced by lentiviral vectors. We succeeded in the generation of murine and importantly also of human endothelial cell lines. The key feature of the established cell lines is that their proliferation status can be strictly controlled while the expression of relevant markers is maintained. This strict control of proliferation was observed in cell clones and in cell pools and was even maintained when two independent immortalizing genes were simultaneously employed. Thus, this strategy is flexible, easy to handle, and reliable. Most importantly, it allows expansion of human cells with a primary-like phenotype.