Embryogenesis is a sequential process of differential gene expression dictated by the epigenetic environment. Exploiting epigenetic influence on phenotypic outcome, biotechnology platforms are developed for reversal of differentiation to achieve genetic reprogramming of adult sources

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Embryogenesis is a sequential process of differential gene expression dictated by the epigenetic environment. Exploiting epigenetic influence on phenotypic outcome, biotechnology platforms are developed for reversal of differentiation to achieve genetic reprogramming of adult sources back to an embryonic state. Such platforms include “therapeutic cloning” and “nuclear reprogramming” that bypass the need for embryo extraction to generate pluripotent stem cell phenotypes from autologous sources. Reprogramming of adult cells to generate customized embryonic-like stem cells offers the future for patient-specific regenerative therapies.
1. induced Pluripotent Stem Cells (iPSC) via nuclear reprogramming
Nuclear reprogramming of adults cells, namely skin fibroblasts, through ectopic introduction of a small number of pluripotency-associated transcription factors is an approach to induce an embryonic stem cell-like phenotype. In the mouse, such an approach has yielded induced pluripotent stem (iPS) cells sufficient for de novo embryogenesis and germline transmission. In humans, transcription factors associated with the pluripotency state were ranked through bioinformatic prioritization and further screened for reprogramming effectiveness through combinatorial viral delivery. Transcription factor sets Oct4, Sox2, c-Myc, and Klf4 or alternatively Oct4, Sox2, Nanog, and Lin28, are sufficient to reprogram human somatic cells to pluripotent stem cells that exhibit the essential characteristics of ESC, including maintenance of the developmental potential to differentiate into advanced derivatives of all three germ layers.
Moreover, iPS-based technology will facilitate the production of cell line panels that closely reflect the genetic diversity of a population enabling the discovery, development, and validation of therapies tailored for each individual. Alleviating technical limitations, including mutation through viral integration, incorporation of oncogenic genes, and establishing reliable patient-specific differentiation protocols are significant advancements that will facilitate clinical translation.
2. therapeutic cloning
Somatic cell nuclear transfer (SCNT) allows transacting factors present within the mammalian oocytes, conserved across species, to reprogram somatic cell nuclei to an undifferentiated state. Therapeutic cloning refers to SCNT in which the nuclear content of a somatic cell from an individual is transferred into an enucleated donor egg to derive blastocysts that contain pluripotent embryonic-like stem cells. In this way, SCNT has produced cloned ESCs from multiple mammalian somatic cell biopsies. The pluripotency of derived cells has been confirmed through germline transmission and reproductive cloning. However, due to technological limitations cloned human blastocysts have only recently been achieved although in low efficiency, and successful isolation of ESCs from the inner cell mass has yet to be demonstrated with human protocols.
Author: dragon web profit

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