Use of RNA for reprogramming somatic cells
Methods for effecting the de-differentiation of somatic cells to cells having stem cell characteristics, in particular pluripotency, include the steps of introducing RNA encoding factors inducing the de-differentiation of somatic cells into the somatic cells and culturing the somatic cells allowing the cells to de-differentiate.
1. A cell population which comprises (i) RNA-transfected somatic cells subjected to electroporation, the RNA-transfected somatic cells containing in vitro transcribed mRNA encoding OCT4, SOX2, KLF4 and c-MYC, wherein the mRNA comprises a 5′-Cap structure and wherein the RNA-transfected cells express alkaline phosphatase and endogenous OCT4, and (ii) somatic cells not subjected to electroporation.
2. The cell population in accordance with claim 1 , wherein the RNA-transfected somatic cells are human cells transfected with RNA.
3. The cell population in accordance with claim 2 , wherein the human cells are adult human dermal fibroblasts transfected with RNA.
4. The cell population in accordance with claim 1 , wherein the somatic cells subjected to electroporation are fibroblasts.
5. The cell population in accordance with claim 1 , wherein the RNA-transfected somatic cells express endogenous OCT4 for at least 7 days.
6. The cell population in accordance with claim 1 , wherein the RNA-transfected somatic cells are fibroblasts deposited at American Type Culture Collection under Catalog No. CCL-186 and subsequently transfected with RNA.
7. The cell population in accordance with claim 1 , wherein the RNA-transfected somatic cells are fibroblasts deposited at American Type Culture Collection under Catalog No. CRL-2097 and subsequently transfected with RNA.
8. The cell population in accordance with claim 1 , wherein the RNA-transfected somatic cells are derived by RNA-transfection of fibroblasts deposited at American Type Culture Collection under Catalog No. CCL-186.
9. The cell population in accordance with claim 1 , wherein the RNA-transfected somatic cells are derived by RNA-transfection of fibroblasts deposited at American Type Culture Collection under Catalog No. CRL-2097.