Methods and compositions for improving the developmental ability and quality of pre-implantation embryos

摘要

A method of improving the developmental ability and quality of in vitro cultured pre-implantation non-human mammalian embryos comprises the steps of: a) Culturing the pre-implantation embryos in vitro to the pre-compaction or pre-blastulation stage; b) Reducing the normal capacity of the embryos to generate ATP by oxidative phosphorylation either by partially reducing the oxygen content of the culture atmosphere or by use of one or more selective oxidative phosphorylation inhibitors and/or uncouplers; c) Culturing the embryos to the compaction or blastocyst stage; and d) Assessing the developmental ability and quality of the embryos for suitability for implantation. A method of improving the viability of an in vitro pure culture of non-human mammalian embryonic-derived cells comprises the steps of: a) Culturing the cells in vitro; b) Reducing the normal capacity of the cells to generate ATP by oxidative phosphorylation either by partially reducing the oxygen content of the culture atmosphere or by use of one or more selective oxidative phosphorylation inhibitors and/or uncouplers; c) Assessing the viability of the cells for use in nuclear transfer. Also described is a culture for improving the developmental and quality of pre-implantation mammalian embryos comprising at least one oxidative phosphorylation inhibitor and/or oxidative phosphorylation uncoupler, in combination with an animal mammalian embryo maintaining medium. The inhibitors is preferably selected from the group consisting of sodium azide, cyanide, rotenone and antimycin A, and any other group of compounds which act as inhibitors of oxidative phosphorylation, and the oxidative phosphorylation uncoupler is preferably selected from the group consisting of 2,4-dinitrophenol (DNP), carbonylcyanide p-trifluoro-methoxyphenylhydrazone (FCCP). The inhibitor and/or uncoupler are present in the culture medium at a concentration of from 0.001 to 1000mM.