Compositions and methods for enhancing bioenergetic status in female germ cells
Compositions and methods comprising bioenergetic agents for restoring the quality of aged oocytes, enhancing oogonial stem cells or improving derivatives thereof (e.g., cytoplasm or isolated mitochondria) for use in fertility-enhancing procedures, are described.
1. A method of culturing a mammalian oocyte, a mammalian oogonial stem cell (OSC), the progeny of a mammalian OSC or a mammalian preimplantation zygote, wherein the OSC is obtained from ovarian tissue, is an isolated non-embryonic stem cell, and is mitotically competent and expresses Vasa, Oct-4, Dazl and Stella and, optionally, a stage-specific embryonic antigen, said method comprising:
culturing said oocyte, OSC, progeny of an OSC or preimplantation zygote in a medium comprising a CD38 inhibitor and a nicotinamide adenine dinucleotide (NAD) precursor,
wherein the CD38 inhibitor is a compound selected from the group consisting of apigenin, luteolin, tyrphostin-8, berberine and SRT-1720,
wherein the NAD precursor is a compound selected from the group consisting of nicotinamide, mononucleotide, nicotinamide riboside or nicotinic acid, and
wherein the CD38 inhibitor and the NAD precursor are present in the medium in an amount effective to enhance the bioenergetic status of said oocyte, OSC, progeny of an OSC or preimplantation zygote cultured in said medium.
2. The method of claim 1 , wherein the medium is selected from the group consisting of cell culture medium, oocyte retrieval solution, oocyte washing solution, oocyte in vitro maturation medium, ovarian follicle in vitro maturation medium, oocyte in vitro fertilization medium, vitrification solution and cryopreservation solution.
3. The method of claim 1 , wherein the NAD precursor is nicotinamide riboside.
4. The method of claim 1 , wherein the NAD precursor is nicotinic acid.
5. The method of claim 1 , wherein a mammalian OSC is cultured.
6. The method of claim 1 , wherein the mammalian oocyte, or OSC, progeny of an OSC or preimplantation zygote is from a human female.
7. The method of claim 6 , wherein the human female is selected from the group consisting of females of advanced maternal age, females suffering from oocyte-related infertility and females with low ovarian reserve.
8. The method of claim 1 , wherein the CD38 inhibitor and the NAD precursor are present in the medium in an amount effective to increase the number of functional mitochondria in the mammalian oocyte, OSC, progeny of an OSC or preimplantation zygote cultured in said medium.
9. The method of claim 1 , wherein the CD38 inhibitor and the NAD precursor are present in the medium in an amount effective to increase the mitochondrial energy of the mammalian oocyte, OSC, progeny of an OSC or preimplantation zygote cultured in said medium.
10. The method of claim 1 , wherein the CD38 inhibitor and the NAD precursor are present in the medium in an amount effective to increase the cellular energy of the mammalian oocyte, OSC, progeny of an OSC or preimplantation zygote cultured in said medium.
11. The method of claim 1 , wherein the CD38 inhibitor is present at a concentration ≧25 μM.
12. The method of claim 1 , wherein the NAD precursor is present at a concentration ≧100 μM.
13. The method of claim 1 , wherein the medium further comprises ovarian tissue, ovarian follicles, bone marrow, umbilical cord blood or peripheral blood.
14. The method of claim 1 , wherein the mammalian OSC expresses a stage-specific embryonic antigen.