Methods and compositions to increase human somatic cell nuclear transfer (scnt) efficiency by removing histone h3-lysine trimethylation, and derivation of human nt-esc

A method for increasing the efficiency of human somatic cell nuclear transfer (SCNT)

• comprising at least one of:
• (iv) contacting a donor human somatic cell or a recipient human oocyte with at least one agent which decreases H3K9me3 methylation in the donor human somatic cell or the recipient human oocyte, wherein the recipient human oocyte is a nucleated or enucleated oocyte;
• enucleating the recipient human oocyte if the human oocyte is nucleated;
• transferring the nuclei from the donor human somatic cell to the enucleated occyte to form a hybrid oocyte;
• and activating the hybrid oocyte to form a human SCNT embryo;
• or
• (v) contacting a hybrid oocyte with at least one agent which decreases H3K9me3 methylation in the hybrid oocyte, where the hybrid oocyte is an enucleated human oocyte comprising the genetic material of a human somatic cell, and activating the hybrid oocyte to form a human SCNT embryo;
• or
• (vi) contacting a human SCNT embryo after activation with at least one agent which decreases H3K9me3 methylation in the human SCNT embryo, wherein the SCNT embryo is generated from the fusion of an enucleated human oocyte with the genetic material of a human somatic cell;
• wherein the decrease of H3K9me3 methylation in any one of the donor human somatic cell, recipient human oocyte, hybrid oocyte or the human SCNT embryo increases the efficiency of the SCNT.

A method for producing a human nuclear transfer embryonic stem cell (hNT-ESC), comprising;

• a. at least one of:
• (i) contacting a donor human somatic cell or a recipient human oocyte with at least one agent which decreases H3K9me3 methylation in the donor human somatic cell or the recipient human oocyte;
• wherein the recipient human oocyte is a nucleated or enucleated oocyte;
• enucleating the recipient human oocyte if the human oocyte is nucleated;
• transferring the nuclei from the donor human somatic cell to the enucleated oocyte to form a hybrid oocyte;
• and
• activating the hybrid oocyte to form a human SCNT embryo;
• or
• (ii) contacting a hybrid oocyte with at least one agent which decreases
• H3K9me3 methylation in the hybrid oocyte, where the hybrid oocyte is an enucleated human oocyte comprising the genetic material of a human somatic cell, and activating the hybrid oocyte to form a human SCNT embryo;
• or
• (iii) contacting a human SCNT embryo after activation with at least one agent
• which decreases H3K9me3 methylation in the SCNT embryo, wherein the SCNT embryo is generated from the fusion of an enucleated human oocyte with the genetic material of a human somatic cell;
• b. incubating the SCNT embryo for a sufficient amount of time to form a blastocyst;
• and collecting at least one blastomere from the blastocyst and culturing the at least one blastomere to form at least one human NT-ESC.