Multifactorial analysis of conditional reprogramming of human keratinocytes

Segni B. Ligaba, Anikita Khurana, Garrett Graham, Ewa Krawczyk, Sandra Jablonski, Emanuel F. Petricoin, Robert I. Glazer, Geeta Upadhyay

Research output: Contribution to journalArticlepeer-review

15 Scopus citations


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, re-verse-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-β 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.

Original languageEnglish
Article numbere0116755
JournalPLoS ONE
Issue number2
StatePublished - 25 Feb 2015
Externally publishedYes


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