Isolated Populations of Female Germline Stem Cells and Cell Preparations and Compositions Thereof

Abstract

The present invention relates to female germline stem cells and their progenitors, methods of isolation thereof, and methods of use thereof.

We claim: 
     
         1 . A method of in vitro fertilization of a female subject, said method comprising the steps of:
 a) producing an oocyte by culturing an isolated cell that is mitotically competent and expresses Vasa, Oct-4, Dazl, Stella and optionally, a stage-specific embryonic antigen, in the presence of an agent that differentiates the cell into an oocyte;   b) fertilizing the oocyte in vitro to form a zygote; and   c) implanting the zygote into the uterus of a female subject.   
     
     
         2 . The method of  claim 1 , wherein the cell can produce oocytes after a duration of at least 1 week, 1 to about 2 weeks, about 2 to about 3 weeks, about 3 to about 4 weeks or more than about 5 weeks post transplantation into a host. 
     
     
         3 . The method of  claim 1 , wherein the cell can produce oocytes after a duration of less than 1 week post transplantation into a host. 
     
     
         4 . The method of  claim 1 , wherein the cell can produce oocytes after a duration of less than about 24 to about 48 hours post transplantation. 
     
     
         5 . The method of  claim 2 , wherein the cell is a female germline stem cell. 
     
     
         6 . The method of  claim 3 , wherein the cell is a female germline stem cell progenitor. 
     
     
         7 . The method of  claim 1 , wherein the cell is a mammalian cell. 
     
     
         8 . The method of  claim 1 , wherein the cell is a human cell. 
     
     
         9 . The method of  claim 1 , wherein the cell is a non-embryonic cell. 
     
     
         10 . The method of  claim 1 , wherein the stage-specific embryonic antigen is stage-specific embryonic antigen-1. 
     
     
         11 . The method of  claim 1 , wherein the cell expresses the stage-specific embryonic antigen. 
     
     
         12 . The method of  claim 11 , wherein the cell is a female germline stem cell progenitor. 
     
     
         13 . The method of  claim 11 , wherein the stage-specific embryonic antigen is stage-specific embryonic antigen-1. 
     
     
         14 . A method of oocyte production, comprising culturing an isolated cell that is mitotically competent and expresses Vasa, Oct-4, Dazl, Stella and optionally, a stage-specific embryonic antigen, in the presence of an agent that differentiates the cell into an oocyte, thereby producing an oocyte. 
     
     
         15 . The method of  claim 15 , wherein the agent is selected from the group consisting of a transforming growth factor, bone morphogenic protein, Wnt family protein, kit-ligand, leukemia inhibitory factor, meiosis-activating sterol, modulator of Id protein function and modulator of Snail/Slug transcription factor function. 
     
     
         16 . A pharmaceutical composition comprising a purified population of cells that are mitotically competent and express Vasa, Oct-4, Dazl, Stella and optionally, a stage-specific embryonic antigen and a pharmaceutically acceptable carrier. 
     
     
         17 . A method of repairing damaged ovarian tissue, comprising providing a therapeutically effective amount of the pharmaceutical composition of  claim 16  to the tissue, wherein the cells engraft into the tissue and differentiate into oocytes, thereby repairing the damaged tissue. 
     
     
         18 . A method of restoring ovarian function in a menopausal female subject, comprising administering a therapeutically effective amount of the pharmaceutical composition of  claim 17  to the subject, wherein the cells engraft into the ovary and differentiate into oocytes, thereby restoring ovarian function in the subject. 
     
     
         19 . A method for oocyte production in a subject, comprising contacting female germline stem cells, or their progenitor cells, of the subject with an agent that differentiates the female germline stem cells, or their progenitor cells, into oocytes, thereby producing oocytes in the subject. 
     
     
         20 . A kit for oocyte production comprising an agent selected from the group consisting of a transforming growth factor, bone morphogenic protein, Wnt family protein, kit-ligand, leukemia inhibitory factor, meiosis-activating sterol, modulator of Id protein function and modulator of Snail/Slug transcription factor function, and instructions for using the agent to differentiate the female germline stem cells, or their progenitor cells, into oocytes, thereby producing oocytes. 
     
     
         21 . A method of expanding female germline stem cells, or their progenitor cells, in vivo, ex vivo or in vitro, comprising contacting the female germline stem cells, or their progenitor cells, with an agent that increases the amount of female germline stem cells, or their progenitor cells, thereby expanding the female germline stem cells, or their progenitor cells. 
     
     
         22 . A kit for expanding female germline stem cells, or their progenitor cells, comprising an agent selected from the group consisting of an insulin-like growth factor, transforming growth factor, bone morphogenic protein, Wnt protein, fibroblast growth factor, sphingosine-1-phosphate, retinoic acid, inhibitor of glycogen synthase kinase-3, Bax inhibitor, caspase inhibitor, inhibitor of nitric oxide production and inhibitor of histone deacetylase activity, and instructions for using the agent to increase the amount of female germline stem cells or their progenitor cells, thereby expanding the female germline stem cells, or their progenitor cells. 
     
     
         23 . A method for oocyte production in a subject, comprising contacting ovarian tissue of the subject with an agent that increases the amount of female germline stem cells, or their progenitor cells, thereby producing oocytes in the subject. 
     
     
         24 . A method of treating infertility in a female subject in need thereof comprising contacting ovarian tissue of the subject with an agent that increases the differentiation of female germline stem cells, or progenitor cells derived from female germline stem cells, into oocytes, thereby treating infertility in the subject. 
     
     
         25 . A method of treating infertility in a female subject in need thereof comprising contacting ovarian tissue of the subject with an agent that increases the amount of female germline stem cells, or their progenitor cells, thereby treating infertility in the subject. 
     
     
         26 . A method of restoring ovarian function in a post-menopausal female subject comprising contacting ovarian tissue of the subject with an agent that increases the amount of female germline stem cells, or their progenitor cells, thereby restoring ovarian function in the subject. 
     
     
         27 . A method of reducing the amount of female germline stem cells, or their progenitor cells, in a subject comprising contacting female germline stem cells, or their progenitor cells, in the subject with an agent that reduces cell proliferation, thereby reducing the amount of female germline stem cells, or their progenitor cells, in the subject. 
     
     
         28 . A method of reducing the amount of female germline stem cells, or their progenitor cells, in a subject comprising contacting female germline stem cells, or their progenitor cells, in the subject with an agent that inhibits cell survival, thereby reducing the amount of female germline stem cells, or their progenitor cells, in the subject. 
     
     
         29 . A kit for reducing the amount of female germline stem cells, or their progenitor cells, comprising an agent that inhibits survival selected from the group consisting of a pro-apoptotic tumor necrosis factor super family member, antagonist of pro-survival Bcl-2 family member function and ceramide, and instructions for using the agent to inhibit cell survival of female germline stem cells, or their progenitors, thereby reducing the amount of female germline stem cells, or their progenitor cells. 
     
     
         30 . A method of reducing the amount of female germline stem cells, or their progenitor cells, in a subject comprising contacting female germline stem cells, or their progenitor cells, in the subject with an agent that promotes cell death, thereby reducing the amount of female germline stem cells, or their progenitor cells, in the subject. 
     
     
         31 . A kit for reducing the amount of female germline stem cells, or their progenitor cells, comprising an agent that promotes cell death selected from the group consisting of a pro-apoptotic tumor necrosis factor superfamily member, agonist of pro-apoptotic Bcl-2 family member function and ceramide, and instructions for using the agent to promote cell death of female germline stem cells, or their progenitor cells, thereby reducing the amount of female germline stem cells, or their progenitor cells. 
     
     
         32 . A kit for contraception in a female subject comprising an agent that decreases the amount of female germline stem cells, or their progenitors cells, and instructions for using the agent to decrease the amount of female germline stem cells, or their progenitor cells, thereby providing contraception to the subject.