TY - JOUR
T1 - Synthetic gene regulation circuits for control of cell expansion
AU - May, Tobias
AU - Butueva, Milada
AU - Bantner, Sara
AU - Markusic, David
AU - Seppen, Jurgen
AU - MacLeod, Roderick A.F.
AU - Weich, Herbert
AU - Hauser, Hansjörg
AU - Wirth, Dagmar
N1 - Copyright: Copyright 2014 Elsevier B.V., All rights reserved.
PY - 2010/2/1
Y1 - 2010/2/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=77049100733&partnerID=8YFLogxK
U2 - https://doi.org/10.1089/ten.tea.2009.0184
DO - https://doi.org/10.1089/ten.tea.2009.0184
M3 - Article
C2 - 19705962
SN - 1937-3341
VL - 16
SP - 441
EP - 452
JO - Tissue Engineering - Part A
JF - Tissue Engineering - Part A
IS - 2
ER -