Human Embryonic Stem Cells Derived by Somatic Cell Nuclear Transfer

Masahito Tachibana,1 Paula Amato,2 Michelle Sparman,1 Nuria Marti Gutierrez,1 Rebecca Tippner-Hedges,1 Hong Ma,1 Eunju Kang,1 Alimujiang Fulati,1 Hyo-Sang Lee,1,6 Hathaitip Sritanaudomchai,3 Keith Masterson,2 Janine Larson,2 Deborah Eaton,2 Karen Sadler-Fredd,2 David Battaglia,2 David Lee,2 Diana Wu,2 Jeffrey Jensen,1,4 Phillip Patton,2 Sumita Gokhale,5 Richard L. Stouffer,1,2 Don Wolf,1 and Shoukhrat Mitalipov1,2,*

Publication is being  scheduled for June 6, 2014 issue of Cell with an early online release date of May 15, 2014.

*Correspondence: mitalipo@ohsu.edu

Summary

Reprogramming somatic cells into pluripotent embryonic stem cells (ESCs) by somatic cell nuclear transfer (SCNT) has been envisioned as an approach for generating patient-matched nuclear transfer (NT)-ESCs for studies of disease mechanisms and for developing specific therapies. Past attempts to produce human NT-ESCs have failed secondary to early embryonic arrest of SCNT embryos. Here, we identified premature exit from meiosis in human oocytes and suboptimal activation as key factors that are responsible for these outcomes. Optimized SCNT approaches designed to circumvent these limitations allowed derivation of human NT-ESCs. When applied to premium quality human oocytes, NT-ESC lines were derived from as few as two oocytes. NT-ESCs displayed normal diploid karyotypes and inherited their nuclear genome exclusively from parental somatic cells. Gene expression and differentiation profiles in human NT-ESCs were similar to embryo-derived ESCs, suggesting efficient reprogramming of somatic cells to a pluripotent state. The human oocyte/embryo research was supported by OHSU institutional funds and by the grant from Leducq Foundation.The nonhuman primate studies were supported by grants from the National Institutes of Health HD063276, HD057121, HD059946, EY021214, and 8P51OD011092.