Question

What is the rate-limiting event in IPS Cell generation process, How this could be overcome? Do you have any alternative idea to get IPS Cells, which could be easy, cheap and faster?

Answer 1

Induced pluripotent stem cells (iPSCs) is a reprogramming technique which from somatic cells to develope newones by the ectopic expression of defined embreyonic transcription factors.Here 4 specific genes (Myc,Oct3/4,Sox2,Klf4)encoding transcription factors could convert somatic cell in to pluripotent stem cells.pluripotent cells hold promise in the field of regenerative medicines,having propagate indefinetely,and give rise to other body parts such as heart,pancreas,liver etc..

iPSs offer a unique chance to model human disease and are already being used to make new discoveries about premature aging,heart disease,cancer and more.

iPCSs can be derived by introducing products of specific sets of pluripotency associated genes or reprogramming factors in to a given cell type(which mentioned above).This combination is most conventional in the formation of iPSCs.Each of the factors can be functionally replaced by related transcription factors,mRNA ,small molecules etc.

It takes slowly for 1-2 weeks for mouse cells and 3-4 for humen.

The fastest way to develope ips cell through removal of reprogramming barriers could enable adult stem/progenitor cells to be reprogrammed into ipsc without ectopic expression of exogeneous factors relying on the endogeneous expression of pluripotency master regulators and appropriate extracellular.

A large number of reprogramming barriers have been include p53-p21 pathway,TGF-Beta and hippo signalling pathways,histone H3 lysine 79methylation,histone deacetylation and MBD3/NuRD complex etc.Depletion or inhibition  of these barriers has been successful to enhance reprogramming efficency of somatic cells.

For evalating the applicable induction of pluripotency in adult stem/progenitor cells without overexpression of exogeneous reprogramming factors,and the chemical or biological molecules which can inhibit barriers or activate enhancers either separately or together are administered in cultures of adult stem cells.Furthermore it is recommended that the cells are cultured in pluripotent supportive reprogramming meadia.

1.isolate and culture donor cells 2.Transduce stem cells associated genes into the cell by viral vectors. 3.harvest and culture the cells 4. iPS cell generate ES like colonies.