Methods and compositions for producing germ cells from bone marrow derived germline stem cells

Abstract

The present invention relates to the use of bone marrow derived germline stem cells and their progenitors, methods of isolation thereof, and methods of use thereof.

1 . An isolated bone marrow cell that is mitotically competent, has an XX karyotype and expresses Vasa, Oct-4, Dazl, Stella, Fragilis and optionally, Nobox, c-Kit and Sca-1. 
     
     
         2 . The isolated cell 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 isolated cell of  claim 1 , wherein the cell can produce oocytes after a duration of less than 1 week post transplantation into a host. 
     
     
         4 . The isolated cell of  claim 1 , wherein the cell can produce oocytes after a duration of less than about 24 to about 48 hours post transplantation into a host. 
     
     
         5 . The isolated cell of  claim 2 , wherein the cell is a bone marrow derived female germline stem cell. 
     
     
         6 . The isolated cell of  claim 3 , wherein the cell is a bone marrow derived female germline stem cell progenitor. 
     
     
         7 . The isolated cell of  claim 1 , wherein the cell is a mammalian cell. 
     
     
         8 . The isolated cell of  claim 1 , wherein the cell is a human cell. 
     
     
         9 . The isolated cell of  claim 1 , wherein the cell is a non-embryonic cell. 
     
     
         10 . The isolated cell of  claim 1 , wherein the cell expresses Nobox. 
     
     
         11 . The isolated cell of  claim 1 , wherein the cell expresses c-Kit. 
     
     
         12 . The isolated cell of  claim 1 , wherein the cell expresses Sca-1. 
     
     
         13 . A method of in vitro fertilization of a female subject, said method comprising the steps of:
 a) producing an oocyte by culturing the isolated cell of  claim 1  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.   
     
     
         14 . A method of oocyte production, comprising culturing the isolated cell of  claim 1  in the presence of an agent that differentiates the cell into an oocyte, thereby producing an oocyte. 
     
     
         15 . The method of  claim 14 , 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, have an XX karyotype and express Vasa, Oct-4, Dazl, Stella, Fragilis and optionally, Nobox, c-Kit and Sca-1 and a pharmaceutically acceptable carrier. 
     
     
         17 . The pharmaceutical composition of  claim 16 , wherein the cells are purified from the bone marrow. 
     
     
         18 . The pharmaceutical composition of  claim 16 , wherein the cells are mammalian cells. 
     
     
         19 . The pharmaceutical composition of  claim 16 , wherein the cells are human cells. 
     
     
         20 . The pharmaceutical composition of  claim 16 , wherein the purified population of cells is about 50 to about 55%, about 55 to about 60%, about 65 to about 70%, about 70 to about 75%, about 75 to about 80%, about 80 to about 85%, about 85 to about 90%, about 90 to about 95% or about 95 to about 100% of the cells in the composition. 
     
     
         21 - 69 . (canceled)