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Multifactorial analysis of conditional reprogramming of human keratinocytes.


ABSTRACT: Co-culture of human primary epithelial cells with irradiated 3T3 fibroblast feeder cells (J2 cells) and the Rho kinase inhibitor Y-27632 (Y) allows for the unrestricted growth of cells of epithelial origin by the process termed conditional reprogramming. To better understand the nature of the signaling processes associated with conditionally reprogrammed cells, the effect of the two critical components of the co-culture conditions, J2 cells and Y, on the growth of human foreskin keratinocytes (HFKs) was evaluated by gene expression profiling, reverse-phase protein arrays and siRNA screening. J2 cells and Y acted cooperatively to down-regulate differentiation, and upregulate proliferation and cell adhesion, including increased pT308Akt and pERK, and reduced TGF-? pathway signaling. These findings establish a mechanistic basis for the unlimited growth potential of human epithelial cells that will be invaluable to assess the effect of genetic changes in pathologic tissues and their response to therapeutic agents.

SUBMITTER: Ligaba SB 

PROVIDER: S-EPMC4340869 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Multifactorial analysis of conditional reprogramming of human keratinocytes.

Ligaba Segni B SB   Khurana Anikita A   Graham Garrett G   Krawczyk Ewa E   Jablonski Sandra S   Petricoin Emanuel F EF   Glazer Robert I RI   Upadhyay Geeta G  

PloS one 20150225 2


Co-culture of human primary epithelial cells with irradiated 3T3 fibroblast feeder cells (J2 cells) and the Rho kinase inhibitor Y-27632 (Y) allows for the unrestricted growth of cells of epithelial origin by the process termed conditional reprogramming. To better understand the nature of the signaling processes associated with conditionally reprogrammed cells, the effect of the two critical components of the co-culture conditions, J2 cells and Y, on the growth of human foreskin keratinocytes (H  ...[more]

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