Oligodendrocyte precursor cell composition and methods of use
The present invention provides a cell culture enriched for sphingolipid enhances neural stem cells (SENSe), particularly oligodendrocyte precursor cells (ODPCs), that do not form teratomas after transplanted in vivo. Methods for producing and use of the invention ODPCs or the cell culture enriched with these ODPCs for stem cell therapy are also provided. The invention method comprises culturing a stem cell culture with a cell culture medium comprising a ceramide compound and a S1P receptor agonist in sequence, overlapping intervals or concurrent manners. The present invention further provides a cellular or gene therapy using a composition comprising a ceramide compound in conjunction with a S1P 1 agonist to proliferate or differentiate endogeneous neural stem cells to ODPCs and further to oligodendrocytes.
1. A cell culture comprising a cell culture medium comprising a ceramide compound in combination with a S1P receptor agonist, wherein said cell culture is enriched with oligodendrocyte precursor cells (ODPCs) capable of further differentiating into oligodendrocytes in vivo without forming teratomas, wherein said ceramide compound is N-(2-hydroxy-1-(hydroxymethyl)ethyl)-oleoylamide (S18), and wherein said S1P receptor agonist is selected from the group consisting of 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol (FTY720), (S)-phosphoric acid mono-[2-amino-3-(4-octyl-phenylamino)-propyl]ester (VPC24191), and a tetraaromatic compound SEW2871.
2. The cell culture of claim 1 , wherein the ODPCs have stellate morphology similar to endogeneous neural stem cells found in adult brain, and do not express Oct-4 marker, but express neural stem cell markers selected from the group consisting of nestin, Sox2, sphingosine kinase 2 (SK2), the S1P receptor 1 (S1P 1 or Edg1), EGF receptor (EGFR), and cyclic nucleotide phosphatase (CNPase).
3. The cell culture of claim 1 , wherein the ODPCs further express oligodendrocyte precursor markers selected from the group consisting of A2B5, GFAP, and NG2 proteoglycan.
4. The cell culture of claim 1 , wherein the ODPCs can further be differentiated to oligodendrocytes which express oligodendrocyte differentiation markers selected from the group consisting of O4, GFP, and myelin basic protein (MBP).
5. The cell culture of claim 1 , wherein said S18 ceramide compound is in a concentration of approximately 1 μM to approximately 500 μM.
6. The cell culture of claim 1 , wherein the S1P receptor agonist is in a concentration of approximately 10 nM to approximately 500 nM.
7. The cell culture of claim 1 , wherein said S1P receptor agonist is 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol (FTY720).
8. The cell culture of claim 1 , wherein said ODPCs are derived from a differentiated stem cell that has been in contact with said cell culture medium for approximately 6 hours to approximately 10 days.
9. The cell culture of claim 8 , wherein said stem cell is an embryonic or adult-derived stem cell.
10. A cell culture enriched with oligodendrocyte precursor cells (ODPCs) capable of further differentiating into oligodendrocytes in vivo without forming teratomas, wherein said cell culture is prepared by the process of contacting said cell culture with a cell culture medium comprising a ceramide compound and a S1P receptor agonist, wherein said ceramide compound is N-(2-hydroxy-1-(hydroxymethyl)ethyl)-oleoylamide (“S18”) and wherein said S1P receptor agonist is selected from the group consisting of 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol (FTY720), (S)-phosphoric acid mono-[2-amino-3-(4-octyl-phenylamino)-propyl]ester (VPC24191), and a tetraaromatic compound SEW2871.
11. The cell culture of claim 10 , wherein the ODPCs have stellate morphology similar to endogeneous neural stem cells found in adult brain, and do not express Oct-4 marker, but express neural stem cell markers selected from the group consisting of nestin, Sox2, sphingosine kinase 2 (SK2), the S1P receptor 1 (S1P 1 or Edg1), EGF receptor (EGFR), and cyclic nucleotide phosphatase (CNPase).
12. The cell culture of claim 10 , wherein the ODPCs further express oligodendrocyte precursor markers selected from the group consisting of A2B5, GFAP, and NG2 proteoglycan.
13. The cell culture of claim 10 , wherein the ODPCs can further be differentiated to oligodendrocytes which express oligodendrocyte differentiation markers selected from the group consisting of O4, GFP, and myelin basic protein (MBP).
14. The cell culture of claim 10 , wherein said S18 ceramide compound is in a concentration of approximately 1 μM to approximately 500 μM.
15. The cell culture of claim 10 , wherein the S1P receptor agonist is in a concentration of approximately 10 nM to approximately 500 nM.
16. The cell culture of claim 10 , wherein said S1P receptor agonist is 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol (FTY720).
17. The cell culture of claim 10 , wherein said ODPCs are derived from a differentiated stem cell that has been in contact with the cell culture medium comprising a ceramide compound and a S1P receptor agonist for approximately 6 hours to approximately 10 days.
18. The cell culture of claim 17 , wherein said stem cell is an embryonic or adult-derived stem cell.
19. The cell culture of claim 8 , where the stem cell is selected from the group consisting of pluripotent stem cell, multipotent stem cell, totipotent stem cell, embryonic stem (ES) cell, inducted pluripotent stem cell (iPS) derived from either a pluripotent cell or a non-pluripotent cell, and any stem cell derived from fetal or adult tissue.