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.

1 - 22 . (canceled) 
     
     
         23 . An oocyte exhibiting increased ATP-generation prepared by a method comprising the step of:
 transferring a composition comprising exogenous, autologous, mitochondria into an oocyte of a subject;   wherein the exogenous, autologous mitochondria are isolated from an autologous oogonial stem cell (OSC) of the subject or the progeny of an autologous OSC of the subject;   wherein the oocyte of the subject has diminished mitochondrial activity without the exogenous, autologous, functional mitochondria;   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, autologous, functional mitochondria increase the ATP-generation of the oocyte, thereby converting the subject's oocyte into an oocyte exhibiting increased ATP-generation and successful fertilization, successful embryonic development or both successful fertilization and successful embryonic development, as compared to said subject's oocyte with diminished mitochondrial activity prior to transfer of said exogenous, autologous, functional mitochondria.   
     
     
         24 . An oocyte exhibiting increased ATP-generation prepared by a method comprising the step of:
 transferring a composition comprising a nuclear-free cytoplasmic fraction of exogenous, autologous, mitochondria into an oocyte of a subject;   wherein the exogenous, autologous, functional mitochondria are isolated from an autologous oogonial stem cell (OSC) of the subject or the progeny of an autologous OSC of the subject;   wherein the oocyte of the subject has diminished mitochondrial activity without the exogenous, autologous, functional mitochondria;   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, autologous, functional mitochondria increase the ATP-generation of the oocyte, thereby converting the subject's oocyte into an oocyte exhibiting increased ATP-generation and successful fertilization, successful embryonic development or both successful fertilization and successful embryonic development, as compared to said subject's oocyte with diminished mitochondrial activity prior to transfer of said nuclear-free cytoplasmic fraction of exogenous, autologous, functional mitochondria.   
     
     
         25 . An oocyte exhibiting increased ATP-generation comprising:
 an oocyte of a subject and exogenous, autologous, functional mitochondria isolated from an autologous oogonial stem cell (OSC) of the subject or the progeny of an autologous OSC of the subject;   wherein the oocyte of the subject has diminished mitochondrial activity without the exogenous, autologous, functional mitochondria;   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, autologous, functional mitochondria increase the ATP-generation of the oocyte, thereby converting the subject's oocyte into an oocyte exhibiting increased ATP-generation and successful fertilization, successful embryonic development or both successful fertilization and successful embryonic development, as compared to said subject's oocyte with diminished mitochondrial activity without said exogenous, autologous, functional mitochondria.   
     
     
         26 . An oocyte exhibiting increased ATP-generation comprising:
 an oocyte of a subject and a nuclear-free cytoplasmic fraction of exogenous, autologous, functional mitochondria isolated from an autologous oogonial stem cell (OSC) of the subject or the progeny of an OSC of the subject;   wherein the oocyte of the subject has diminished mitochondrial activity without the exogenous, autologous, functional mitochondria;   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, autologous, functional mitochondria increase the ATP-generation of the oocyte, thereby converting the subject's oocyte into an oocyte exhibiting increased ATP-generation and successful fertilization, successful embryonic development or both successful fertilization and successful embryonic development, as compared to said subject's oocyte with diminished mitochondrial without said exogenous, autologous, functional mitochondria.   
     
     
         27 . The oocyte exhibiting increased ATP-generation of any one of  claim 23  or  24 , wherein the method further comprises obtaining said mitochondria or composition. 
     
     
         28 . The oocyte exhibiting increased ATP-generation any one of  claim 23  or  24 , wherein the mitochondria or composition is transferred into the oocyte by injection. 
     
     
         29 . The oocyte exhibiting increased ATP-generation any one of  claim 23  or  24 , wherein the mitochondria or composition is transferred into the oocyte by intracytoplasmic sperm injection. 
     
     
         30 . The oocyte exhibiting increased ATP-generation any one of  claim 23  or  24 , wherein the mitochondria or composition is transferred substantially simultaneously with intracytoplasmic sperm injection. 
     
     
         31 . The oocyte exhibiting increased ATP-generation of any one of  claims 23 - 26 , wherein the OSC or the progeny of an OSC is obtained from a mammal. 
     
     
         32 . The oocyte exhibiting increased ATP-generation of any one of  claims 23 - 26 , wherein the OSC or the progeny of an OSC is obtained from a human. 
     
     
         33 . The oocyte exhibiting increased ATP-generation of  claim 32 , 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. 
     
     
         34 . The oocyte exhibiting increased ATP-generation of any one of  claims 23 - 26 , wherein the oocyte is used for in vitro fertilization. 
     
     
         35 . The oocyte exhibiting increased ATP-generation of any one of  claims 23 - 26 , wherein the oocyte comprises 1×10 3  to 5×10 4  exogenous, autologous, functional mitochondria. 
     
     
         36 . The oocyte exhibiting increased ATP-generation of any one of  claims 23 - 26 , wherein at least 75% of the exogenous, autologous functional mitochondria are high ATP-generating mitochondria. 
     
     
         37 . The oocyte exhibiting increased ATP-generation of any one of  claims 23 - 26 , wherein less than about 5% to about 25% of the mtDNA of the exogenous, autologous functional mitochondria in the composition comprises a deletion mutation within nucleotides 8470-13447 of the mitochondrial genome.