Methods and Compositions for Producing Germ Cells from Peripheral Blood Derived Germline Stem Cells

Abstract

The application describes the use of peripheral blood derived germline stem cells and their progenitors, methods of isolation thereof, and methods of use thereof.

1 . An isolated peripheral blood cell that is mitotically competent, has an XX karyotype and expresses Vasa, Dazl and Stella. 
     
     
         2 - 9 . (canceled) 
     
     
         10 . 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.   
     
     
         11 . 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. 
     
     
         12 . (canceled) 
     
     
         13 . A pharmaceutical composition comprising a purified population of the isolated cells of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         14 - 17 . (canceled) 
     
     
         18 . A method of oocyte production in a subject, comprising providing the pharmaceutical composition of  claim 13  to a tissue of the subject, wherein the cells engraft into the tissue and differentiate into oocytes, thereby producing oocytes in the subject. 
     
     
         19 . (canceled) 
     
     
         20 . A method of inducing folliculogenesis in a subject, comprising providing the pharmaceutical composition of  claim 13  to the subject, wherein the cells engraft into a tissue of the subject and differentiate into oocytes within follicles, thereby inducing folliculogenesis in the subject. 
     
     
         21 . (canceled) 
     
     
         22 . A method of treating infertility in a female subject in need thereof comprising administering a therapeutically effective amount of the pharmaceutical composition of  claim 13  to the subject, wherein the cells engraft into the ovary and differentiate into oocytes, thereby treating infertility. 
     
     
         23 . A method of repairing damaged ovarian tissue in a subject, comprising providing a therapeutically effective amount of the pharmaceutical composition of  claim 13  to the tissue, wherein the cells engraft into the tissue and differentiate into oocytes, thereby repairing the damaged tissue in the subject. 
     
     
         24 - 26 . (canceled) 
     
     
         27 . A method of restoring ovarian function in a menopausal female subject, comprising administering a therapeutically effective amount of the pharmaceutical composition of  claim 13  to the subject, wherein the cells engraft into the ovary and differentiate into oocytes, thereby restoring ovarian function in the subject. 
     
     
         28 - 31 . (canceled) 
     
     
         32 . A method for oocyte production in a subject, comprising contacting peripheral blood derived female germline stem cells, or their progenitor cells, of the subject with an agent that differentiates the peripheral blood derived female germline stem cells, or their progenitor cells, into oocytes, thereby producing oocytes in the subject. 
     
     
         33 . (canceled) 
     
     
         34 . 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 peripheral blood derived female germline stem cells of  claim 32 , or their progenitor cells, into oocytes, thereby producing oocytes. 
     
     
         35 . A method of expanding peripheral blood derived female germline stem cells, or their progenitor cells, in vivo, ex vivo or in vitro, comprising contacting the peripheral blood derived female germline stem cells of  claim 32 , or their progenitor cells, with an agent that increases the amount of peripheral blood derived female germline stem cells, or their progenitor cells, thereby expanding the peripheral blood derived female germline stem cells, or their progenitor cells. 
     
     
         36 . (canceled) 
     
     
         37 . (canceled) 
     
     
         38 . A kit for expanding peripheral blood derived 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 peripheral blood derived female germline stem cells of  claim 32 , or their progenitor cells, thereby expanding the peripheral blood derived female germline stem cells, or their progenitor cells. 
     
     
         39 . A method for oocyte production in a subject, comprising contacting peripheral blood derived female germline stem cells, or their progenitor cells, of the subject with an agent that increases the amount of peripheral blood derived female germline stem cells of  claim 32 , or their progenitor cells, thereby producing oocytes in the subject. 
     
     
         40 . (canceled) 
     
     
         41 . (canceled) 
     
     
         42 . A method of restoring fertility to a female subject who desires restored fertility, comprising administering a therapeutically effective amount of the peripheral blood derived female germline stem cells of  claim 32 , or their progenitor cells, to the subject, wherein the cells engraft into a tissue and differentiate into oocytes, thereby restoring fertility in the subject. 
     
     
         43 . (canceled) 
     
     
         44 . A method of protecting fertility in a female subject undergoing or expected to undergo chemotherapy, radiotherapy or both treatments, comprising providing an agent that protects against reproductive injury prior to or concurrently with chemotherapy, radiotherapy or both treatments and providing a peripheral blood derived female germline stem cell of  claim 32 , or its progenitor cell, to the subject, wherein the cell engrafts into a tissue and differentiates into an oocyte, thereby protecting fertility in the subject. 
     
     
         45 . (canceled) 
     
     
         46 . A kit for protecting fertility in a female subject undergoing or expected to undergo chemotherapy, radiotherapy or both treatments, comprising an agent of  claim 45  selected from the group consisting of S1P, a Bax antagonist, or any agent that increases SDF-1 activity and instructions for using the agent to protect peripheral blood derived female germline stem cells of  claim 32 , or their progenitor cells, against reproductive injury thereby protecting fertility in the female subject. 
     
     
         47 . A method for in vitro fertilization of a female subject, said method comprising the steps of:
 a) producing an oocyte by contacting a peripheral blood derived female germline stem cell of  claim 32 , or its progenitor cell, with an agent that differentiates the peripheral blood derived female germline stem cell, or its progenitor 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.   
     
     
         48 . An isolated peripheral blood cell that is mitotically competent, has an XY karyotype and expresses Vasa and Dazl. 
     
     
         49 - 52 . (canceled) 
     
     
         53 . A method of restoring or enhancing spermatogenesis, comprising providing a peripheral blood derived male germline stem cell, or its progenitor cell, to the testes of a male subject, wherein the cell engrafts into the seminiferous epithelium and differentiates into a sperm cell, thereby restoring or enhancing spermatogenesis. 
     
     
         54 . A method of restoring fertility to a male subject having undergone chemotherapy or radiotherapy, or both and who desires restored fertility, comprising administering a therapeutically effective amount of the peripheral blood derived male germline stem cells of  claim 53 , or their progenitor cells, to the subject, wherein the cells engraft into the seminiferous epithelium and differentiate into sperm cells, thereby restoring fertility. 
     
     
         55 . A method of reducing the amount of peripheral blood derived germline stem cells, or their progenitor cells, in a subject comprising contacting peripheral blood derived germline stem cells, or their progenitor cells, in the subject with an agent that (i) reduces cell proliferation, (ii) inhibits cell survival, or (iii) promotes cell death, thereby reducing the amount of peripheral blood derived germline stem cells, or their progenitor cells, in the subject. 
     
     
         56 - 59 . (canceled) 
     
     
         61 . A kit for reducing the amount of female germline stem cells, or their progenitor cells, comprising an agent that inhibits survival is selected from the group consisting of a pro-apoptotic tumor necrosis factor, and instructions for using the agent to inhibit cell survival of peripheral blood derived germline stem cells in accordance with the method of  claim 55 . 
     
     
         62 - 65 . (canceled) 
     
     
         66 . A kit for reducing the amount of peripheral blood derived germline stem cells, or their progenitor cells, comprising an agent 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 in accordance with the method of  claim 55 . 
     
     
         67 . (canceled) 
     
     
         68 . (canceled) 
     
     
         69 . A method of providing contraception to a subject comprising contacting peripheral blood derived germline stem cells, or their progenitor cells, with an agent that decreases the amount of peripheral blood derived germline stem cells, or their progenitor cells, thereby providing contraception to the subject. 
     
     
         70 . A kit for contraception in a subject comprising an agent of  claim 69 , and instructions for using the agent to decrease the amount of peripheral blood derived germline stem cells, or their progenitor cells, thereby providing contraception to the subject.