Journal
TISSUE ENGINEERING PART A
Volume 16, Issue 2, Pages 441-452Publisher
MARY ANN LIEBERT, INC
DOI: 10.1089/ten.tea.2009.0184
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Funding
- Deutsche Forschungsgemeinschaft [Wi 2624, SFB 599/D8]
- Bundesministerium fur Bildung und Forschung [FKZ0313940]
- EU [MEST-CT-2004-504990]
- Helmholtz Society
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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.
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