PRODUCTION OF REPROGRAMMED PLURIPOTENT CELLS
The present invention provides a method of producing a reprogrammed cell, said method comprising exposing Stro-1 + multipotential cells and/or progeny cells thereof to one or more potency-determining factors under conditions sufficient to reprogram the cells. The present invention also provides cells produced by such a method and cells differentiated therefrom in addition to various uses of those cells.
1 . A method of producing a reprogrammed cell, said method comprising exposing Stro-1 + multipotential cells and/or progeny cells thereof to one or more potency-determining factors under conditions sufficient to reprogram the cells.
2 . A method of producing a reprogrammed cell, said method comprising exposing a population of cells enriched for Stro-1 + multipotential cells and/or progeny cells thereof to one or more potency-determining factors under conditions sufficient to reprogram the cells.
3 . The method according to claim 1 or 2 , wherein the Stro-1 + multipotential cells and/or progeny cells thereof, or cell populations enriched for Stro-1 + multipotential cells and/or progeny cells thereof, are derived from adipose tissue, dental pulp tissue, or bone marrow.
4 . The method according to claim 1 or 2 additionally comprising culturing the exposed cells to produce reprogrammed cells.
5 . The method according to claim 1 or 2 additionally comprising isolating the reprogrammed cells.
6 . The method according to any one of claims 1 to 5 comprising culturing the exposed cells to obtain reprogrammed cells having broader differentiation potential than the Stro-1 + multipotential cells and/or progeny cells thereof.
7 . The method according to any one of claims 1 to 6 , wherein the one or more potency-determining factors are individually or collectively selected from the group consisting of Oct4, Sox2, Klf4, Nanog, Lin28, c-Myc, bFGF, SCF, TERT, SV40 large T antigen, HPV16E6, HPV16E7, Bmil, Fbx15, Eras, ECAT15-2, Tcl1, β-catenin, ECAT1, ESG1, Dnmt3L, ECAT8, Gdf3, Sox15, ECAT15-1, Fthl17, Sal14, Rex1, UTF1, Stella, Stat3 and Grb2 or a compound having the same or similar activity to one or more of said factors.
8 . The method according to any one of claims 1 to 7 , wherein one or more potency determining factor(s) is a chemical, a peptide, a siRNA, a short hairpin RNA or a microRNA.
9 . The method according to any one of claims 1 to 7 , wherein the one or more potency-determining factors are individually or collectively selected from the group consisting of:
(i) Oct4;
(ii) a combination of Oct4 and Sox2;
(iii) a combination of Oct4, Sox2 and at least one of Nanog and Lin28;
(iv) a combination of Oct4, Klf4 and c-Myc;
(v) a combination of Oct4, Sox2 and Klf4;
(vi) a combination of OCT4, Sox2, Klf4 and c-Myc;
(vii) a combination of Oct4, Sox2, Nanog and Lin28;
(viii) a combination of Oct4, Sox2, Klf4, c-Myc, Nanog and Lin28; and
(ix) any one of (i) to (x) additionally in combination with a chemical, a peptide, a siRNA, a shRNA or a microRNA.
10 . The method according to any one of claims 1 to 9 , wherein the Stro-1 + multipotential cells and/or progeny cells thereof are obtained from a post-natal mammal.
11 . The method according to claim 10 , wherein the mammal is human.
12 . The method according to claim 1 or 2 , wherein exposing the Stro-1 + multipotential cells and/or progeny cells thereof to one or more potency-determining factors comprises introducing nucleic acid comprising a sequence encoding one or more potency-determining factors operably linked to a promoter into the Stro-1 + multipotential cells and/or progeny cells thereof.
13 . The method according to claim 12 , comprising administering a plurality of nucleic acids each comprising a sequence encoding a distinct potency-determining factor operably linked to a promoter.
14 . The method according to claim 12 or 13 , wherein the nucleic acid(s) are within one or more vector(s).
15 . The method according to claim 14 , wherein the vector(s) is(are) a viral vector(s).
16 . The method according to claim 12 , wherein the nucleic acid(s) do(es) not integrate into the genome of the Stro-1 + multipotential cells and/or progeny cells thereof.
17 . The method according to claim 1 or 2 , wherein the reprogrammed cells are pluripotent.
18 . The method according to claim 1 or 2 , wherein the reprogrammed cells (i) express a cell marker selected from the group consisting of Oct-4, Nanog, SSEA3, SSEA4, Tra-1-60 and Tra-1-81; (ii) exhibit morphology characteristic of pluripotent cells; and/or (iii) form teratomas when introduced into an immunocompromised animal.
19 . A cell produced by performing the method according to any one of claims 1 to 18 .
20 . A cell population enriched for pluripotent cells produced by performing the method according to the method of any one of claims 1 to 18 .
21 . The enriched population of cells according to claim 20 , wherein the pluripotent cells account for at least 60% of the population.
22 . A cell or population of cells differentiated from the cell or population according to any one of claims 18 to 21 .
23 . A Stro-1 + multipotential cell and/or progeny cell thereof comprising a nucleic acid encoding a potency determining factor operably linked to a heterologous promoter.
24 . Use of the cell or population according to any one of claims 19 to 23 in medicine.
25 . A method of treating or preventing a disease or disorder, the method comprising administering the cell or population according to any one of claims 19 to 23 to a subject in need thereof.
26 . A method of screening for compounds useful in the treatment or prevention of a disease or disorder, the method comprising exposing the cell or population according to any one of claims 19 to 23 to said compounds.