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Human Somatic Cell Nuclear Transfer Using Adult Cells
Chung, Y.,Eum, J.,Lee, J.,Shim, S.,Sepilian, V.,Hong, S.,Lee, Y.,Treff, Nathan R.,Choi, Y.,Kimbrel, Erin A.,Dittman, Ralph E.,Lanza, R.,Lee, D. Cell Press 2014 Cell stem cell Vol.14 No.6
Derivation of patient-specific human pluripotent stem cells via somatic cell nuclear transfer (SCNT) has the potential for applications in a range of therapeutic contexts. However, successful SCNT with human cells has proved challenging to achieve, and thus far has only been reported with fetal or infant somatic cells. In this study, we describe the application of a recently developed methodology for the generation of human ESCs via SCNT using dermal fibroblasts from 35- and 75-year-old males. Our study therefore demonstrates the applicability of SCNT for adult human cells and supports further investigation of SCNT as a strategy for regenerative medicine.
Chung, Young Gie,Matoba, Shogo,Liu, Yuting,Eum, Jin Hee,Lu, Falong,Jiang, Wei,Lee, Jeoung Eun,Sepilian, Vicken,Cha, Kwang Yul,Lee, Dong Ryul,Zhang, Yi Elsevier 2015 Cell stem cell Vol.17 No.6
<P><B>Summary</B></P> <P>The extremely low efficiency of human embryonic stem cell (hESC) derivation using somatic cell nuclear transfer (SCNT) limits its potential application. Blastocyst formation from human SCNT embryos occurs at a low rate and with only some oocyte donors. We previously showed in mice that reduction of histone H3 lysine 9 trimethylation (H3K9me3) through ectopic expression of the H3K9me3 demethylase Kdm4d greatly improves SCNT embryo development. Here we show that overexpression of a related H3K9me3 demethylase KDM4A improves human SCNT, and that, as in mice, H3K9me3 in the human somatic cell genome is an SCNT reprogramming barrier. Overexpression of KDM4A significantly improves the blastocyst formation rate in human SCNT embryos by facilitating transcriptional reprogramming, allowing efficient derivation of SCNT-derived ESCs using adult Age-related Macular Degeneration (AMD) patient somatic nuclei donors. This conserved mechanistic insight has potential applications for improving SCNT in a variety of contexts, including regenerative medicine.</P> <P><B>Highlights</B></P> <P> <UL> <LI> H3K9me3 is a barrier for human SCNT reprogramming as in mouse </LI> <LI> Injection of human KDM4A mRNA improves SCNT success in mouse oocytes </LI> <LI> KDM4A also improves human SCNT blastocyst formation and NT-ESC derivation </LI> <LI> H3K9me3 removal facilitates zygotic genome activation in human SCNT embryos </LI> </UL> </P> <P><B>Graphical Abstract</B></P> <P>[DISPLAY OMISSION]</P>