Nuclear transfer
AssigneeSung Kwang Medical Foundation
InventorsYoung Gie Chung, Dong Ryul Lee, Jin Hee Eum
Life of this patent
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Abstract
The present invention provides methods and compostions to improve the efficiency of somatic cell nuclear transfer (SCNT). There is increasing evidence that the epigenetic state of donor nuclei has a significant impact on potential of nuclear transfer embryos to develop into blastocysts, from which pluripotent stem cells are derived. Strategic application of histone agents, capable of altering epigenetic state such as methylation, allows zygotic activation and robust blastocyst generation.
Claims12
A method of increasing the generation of a blastocyst, comprising:
- providing an enucleated human oocyte;
- generating a nuclear transferred (NT) oocyte by adding at least one nucleus of at least one human donor cell to the enucleated human oocyte;
- activating the NT oocyte by incubation in an activation medium to generate an activated NT oocyte;
- culturing the activated NT oocyte at the 8-cell stage, in a media comprising CARM1 conjugated to a cell penetrating peptide (CPP) to generate a modified, activated NT oocyte;
- wherein said culturing of the activated NT oocyte increases the chance of generating a blastocyst and the rate of successfully generating a blastocyst from the modified, activated NT oocyte is at least 100% greater when compared to the rate of successfully generating a blastocyst from an unmodified, activated NT oocyte;
- generating a blastocyst from the modified, activated NT oocyte;
- and
- isolating inner cell mass (ICM) cells from the blastocyst, wherein the ICM cells are capable of further culturing as a NT-hPSC cell line.
A method of increasing the generation of a blastocyst comprising:
- removing the host nucleus of a human oocyte to generate an enucleated human oocyte;
- generating a nuclear transferred (NT) oocyte by adding at least one nucleus of at least one human donor cell to the enucleated human oocyte;
- activating the NT oocyte by incubation in an activation medium to generate an activated NT oocyte;
- culturing the activated NT oocyte at the 8-cell stage, in a media comprising CARM1 conjugated to a cell penetrating peptide (CPP) to generate a modified, activated NT oocyte;
- wherein said culturing of the activated NT oocyte increases the chance of generating a blastocyst and the rate of successfully generating a blastocyst from the modified, activated NT oocyte is at least 100% greater when compared to the rate of successfully generating a blastocyst from an unmodified, activated NT oocyte;
- generating a blastocyst from the modified activated NT oocyte;
- and
- isolating inner cell mass (ICM) cells from the blastocyst, wherein the ICM cells are capable of further culturing as a NT-hPSC cell line.
Dependencies are inferred from the claim text.