Human Stem Cell Materials and Methods
Monocyte derived adult stem cells (MDSCs) isolated from peripheral blood of mammals is provided, along with pharmaceutical compositions containing an MDSC, kits containing a pharmaceutical composition, and methods of preparing, propagating and using MDSCs or differentiated derivatives thereof. The uses of these biological materials include methods of treating disorders or diseases, as well as methods of ameliorating a symptom associated with any such disorder or disease.
1 . An isolated monocyte-derived stem cell (MDSC) wherein the cell exhibits a surface antigen selected from the group consisting of MAC-1, CD14, CD34, CD40 and CD45.
2 . An isolated monocyte-derived stem cell (MDSC) wherein the cell produces detectable levels of a cytokine selected from the group consisting of interleukin-1β (IL-1β), interleukin-6 (IL-6) and interleukin-12 p70 (IL-12 p70).
3 . An isolated monocyte-derived stem cell (MDSC) wherein the cell exhibits phagocytic activity.
4 . An isolated monocyte-derived stem cell (MDSC) wherein the cell is resistant to dispersion by an agent selected from the group consisting of trypsin, EDTA, and dispase.
5 . An isolated monocyte-derived stem cell (MDSC) wherein the cell is an adult human cell, further wherein the cell exhibits a surface antigen selected from the group consisting of MAC-1, CD14, CD34, CD40 and C45, further wherein the cell produces a cytokine selected from the group consisting of interleukin-1β (IL-1β), interleukin-6 (IL-6) and interleukin-12 p70 (IL-12 p70), further wherein the cell is resistant to dispersion by an agent selected from the group consisting of trypsin, EDTA, and dispase, and further wherein the cell exhibits phagocytic activity.
6 . A method of preparing an isolated MDSC comprising the steps of:
a) isolating a peripheral-blood monocyte (PBM);
b) contacting said PBM with an effective amount of a mitogenic compound selected from the group consisting of macrophage colony-stimulating factor (M-CSF), interleukin-6 (IL-6), and leukemia inhibitory factor (LIF); and
c) culturing said PBM under conditions suitable for propagation of said cell, thereby obtaining a preparation of an isolated MDSC.
7 . The method according to claim 6 wherein the PPM is cryopreserved prior to contacting said PBM with a mitogenic compound.
8 . The method according to claim 6 further comprising cryopreservation of said MDSC.
9 . The method according to claim 6 wherein the PBM is a mammalian PBM.
10 . The method according to claim 9 wherein the PPM is a human PBM.
11 . The method according to claim 10 wherein the PBM is an adult human PBM.
12 . An isolated MDSC obtained by the method according to claim 6 .
13 . A method of generating a differentiated cell comprising the steps of:
a) isolating a MDSC by the method according to claim 6; and
b) contacting the stem cell with an amount of an inducing agent effective to induce differentiation of the cell, thereby generating a differentiated cell.
14 . The method according to claim 13 further comprising cryopreserving said differentiated cell.
15 . The method according to claim 13 further comprising culturing said differentiated cell.
16 . The method according to claim 15 wherein the differentiated ell/inducing agent are selected from the group consisting of a neuronal cell/nerve growth factor (bNGF), an endothelial cell/-vascular endothelial growth factor (VEGF), an epithelial cell/epidermal growth factor (EGF), a T-lymphocyte/interleukin-2 (IL-2), a macrophage/lipopolysaccharide (LPS), and a hepatocyte/hepatocyte growth factor (HGF).
17 . The method according to claim 13 wherein the MDSC is a human MDSC.
18 . The method according to claim 17 wherein the MDSC is an adult human MDSC.
19 . A method for identifying a cell type-specific therapeutic agent comprising
(a) contacting a first differentiated cell obtained according to the method of claim 13 and a candidate therapeutic agent;
(b) further contacting a second differentiated cell obtained according to the method of claim 13 and the candidate therapeutic agent, wherein the first and second differentiated cells are different cell types; and
(c) measuring the viability of the first differentiated cell relative to the viability of the second differentiated cell, wherein a difference in viabilities identifies the candidate therapeutic agent as a cell type-specific therapeutic agent.
20 . A method of treating a disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of a MDSC.
21 . The use of a MDSC to treat a disorder according to claim 20 wherein the MDSC is isolated from the organism to receive treatment.
22 . The use of the MDSC according to claim 21 wherein the organism is a human.
23 . A method of treating a neuronal cell disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of a neuronal cell obtained by the method according to claim 16 .
24 . A method of treating an endothelial cell disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of an endothelial cell obtained by the method according to claim 16 .
25 . A method of treating an epithelial cell disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of an epithelial cell obtained by the method according to claim 16 .
26 . A method of treating a T-lymphocyte disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of a T-lymphocyte obtained by the method according to claim 16 .
27 . A method of treating a macrophage cell disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of a macrophage obtained by the method according to claim 16 .
28 . A method of treating a hepatocyte disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of an hepatocyte obtained by the method according to claim 16 .
29 . A method of ameliorating a symptom associated with a neuronal cell disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of a neuronal cell obtained by the method according to claim 16 .
30 . A method of ameliorating a symptom associated with an endothelial cell disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of an endothelial cell obtained by the method according to claim 16 .
31 . A method of ameliorating a symptom associated with an epithelial cell disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of an epithelial cell obtained by the method according to claim 16 .
32 . A method of ameliorating a symptom associated with a T-lymphocyte disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of a T-lymphocyte obtained by the method according to claim 16 .
33 . A method of ameliorating a symptom associated with a macrophage cell disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of a macrophage obtained by the method according to claim 16 .
34 . A method of ameliorating a symptom associated with a hepatocyte disorder amenable to cell-based treatment comprising administering a pharmaceutically effective amount of an hepatocyte obtained by the method according to claim 16 .
35 . A pharmaceutical composition comprising a MDSC and a pharmaceutically acceptable diluent, carrier or medium.
36 . A kit comprising the pharmaceutical composition according to claim 35.