Compositions and Methods for Autologous Germline Mitochondrial Energy Transfer

Abstract

Oogonial stem cell (OSC)-derived compositions, such as nuclear free cytoplasm or isolated mitochondria, and uses of OSC-derived compositions in autologous fertility-enhancing procedures are described.

What is claimed is: 
     
         1 . An oocyte with increased ATP-generating capacity comprising: a composition including exogenous functional mitochondria, wherein the exogenous functional mitochondria are obtained from an autologous ovarian oogonial stem cell (OSC) or the progeny of an autologous ovarian OSC, wherein the OSC is a non-embryonic stem cell that is mitotically competent and expresses Vasa, Oct-4, Dazl, Stella, and optionally, a stage-specific embryonic antigen; and wherein the exogenous functional mitochondria increase the ATP-generating capacity of the oocyte as compared to an autologous oocyte lacking the exogenous functional mitochondria. 
     
     
         2 . An oocyte with increased ATP-generating capacity comprising:
 a composition including a nuclear-free cytoplasmic fraction of exogenous, functional mitochondria obtained from an autologous ovarian oogonial stem cell (OSC) or the progeny of an autologous ovarian OSC, wherein the OSC is a non-embryonic stem cell that is mitotically competent and expresses Vasa, Oct-4, Dazl, Stella, and optionally, a stage-specific embryonic antigen; and wherein the exogenous functional mitochondria increase the ATP-generating capacity of the oocyte as compared to an autologous oocyte lacking the exogenous functional mitochondria.   
     
     
         3 . An oocyte with increased ATP-generating capacity prepared by a method comprising the step of transferring into an oocyte a composition comprising one or more exogenous, functional mitochondria obtained from an autologous ovarian oogonial stem cell (OSC) or the progeny of an autologous ovarian OSC, wherein the OSC is a non-embryonic stem cell that is mitotically competent and expresses Vasa, Oct-4, Dazl, Stella, and optionally, a stage-specific embryonic antigen; and wherein the exogenous functional mitochondria increase the ATP-generating capacity of the oocyte as compared to an autologous oocyte lacking the exogenous functional mitochondria. 
     
     
         4 . An oocyte with increased ATP-generating capacity prepared by a method comprising the step of transferring into an oocyte a composition comprising a nuclear-free cytoplasmic fraction of exogenous, functional mitochondria obtained from an autologous ovarian oogonial stem cell (OSC) or the progeny of an autologous ovarian OSC, wherein the OSC is a non-embryonic stem cell that is mitotically competent and expresses Vasa, Oct-4, Dazl, Stella, and optionally, a stage-specific embryonic antigen; and wherein the exogenous functional mitochondria increase the ATP-generating capacity of the oocyte as compared to an autologous oocyte lacking the exogenous functional mitochondria. 
     
     
         5 . The oocyte with increased ATP-generating capacity of  claim 3  or  claim 4 , wherein the method further comprises obtaining said composition. 
     
     
         6 . The oocyte with increased ATP-generating capacity of  claim 3  or  claim 4 , wherein the composition is transferred into the oocyte by injection. 
     
     
         7 . The oocyte with increased ATP-generating capacity of  claim 3  or  claim 4 , wherein the composition is transferred into the oocyte by introcytoplasmic sperm injection. 
     
     
         8 . The oocyte with increased ATP-generating capacity of  claim 3  or  claim 4 , wherein the composition is transferred substantially simultaneously with introcytoplasmic sperm injection. 
     
     
         9 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein the ovarian OSC or the progeny of an ovarian OSC is obtained from a mammal. 
     
     
         10 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein the ovarian OSC or the progeny of an ovarian OSC is obtained from a human. 
     
     
         11 . The oocyte with increased ATP-generating capacity of  claim 10 , wherein the human is a female selected from the group consisting of females of advanced maternal age, females suffering from oocyte-related infertility and females with low ovarian reserve. 
     
     
         12 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein the oocyte is used for in vitro fertilization. 
     
     
         13 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein the composition comprises 1×10 3  to 5×10 4  exogenous functional mitochondria. 
     
     
         14 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein the oocyte with increased ATP-generating capacity comprises 1×10 3  to 5×10 4  more functional mitochondria than an autologous oocyte lacking the exogenous functional mitochondria. 
     
     
         15 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein at least 75% of the exogenous functional mitochondria in the composition are high ATP-generating mitochondria. 
     
     
         16 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein at least 75% of the exogenous functional mitochondria in the oocyte are high ATP-generating mitochondria as compared to an autologous oocyte lacking the exogenous functional mitochondria. 
     
     
         17 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein less than about 5% to about 25% of the mtDNA of the exogenous functional mitochondria in the composition comprises a deletion mutation within nucleotides 8470-13447 of the mitochondrial genome. 
     
     
         18 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein less than about 5% to about 25% of the mtDNA of the exogenous functional mitochondria in the oocyte comprises a deletion mutation within nucleotides 8470-13447 of the mitochondrial genome as compared to an autologous oocyte lacking the exogenous functional mitochondria. 
     
     
         19 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein the OSC or the progeny of an OSC produces at least two fold more, at least three fold more, at least four fold more, or at least ten fold more ATP per fg mtDNA than an ovarian somatic cell. 
     
     
         20 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein the OSC or the progeny of an OSC produces at least two fold more, at least three fold more, at least four fold more, or at least ten fold more ATP per fg mtDNA than a mesenchymal stem cell. 
     
     
         21 . The oocyte with increased ATP-generating capacity of any one of  claims 1 - 5 , wherein the OSC or progeny of an OSC produces greater or equivalent amounts of ATP/cell with approximately 100 fold less mitochondria as compared to ovarian somatic cells. 
     
     
         22 . An oocyte with increased ATP-generating capacity comprising: a composition including exogenous functional mitochondria, wherein the exogenous functional mitochondria are obtained from an autologous ovarian oogonial stem cell (OSC) or the progeny of an autologous ovarian OSC, wherein the autologous OSC or the progeny of an OSC are obtained from a human female selected from the group consisting of females of advanced maternal age, females suffering from oocyte-related infertility and females with low ovarian reserve, wherein the OSC is a non-embryonic stem cell that is mitotically competent and expresses Vasa, Oct-4, Dazl, Stella, and optionally, a stage-specific embryonic antigen; and wherein the exogenous functional mitochondria increase the ATP-generating capacity of the oocyte as compared to an autologous oocyte lacking the exogenous functional mitochondria.