We claim:
1. A method of differentiating progenitor cells, comprising the steps of:
(a) contacting said progenitor cells with a differentiating agent; and
(b) introducing into said progenitor cells a nucleic acid molecule encoding a MEF2 polypeptide or an active fragment thereof,
thereby differentiating said progenitor cells to produce a cell population containing protected neuronal cells.
2. The method of claim 1, wherein said MEF2 polypeptide is human MEF2C, or an active fragment thereof.
3. The method of claim 1, wherein said MEF2 polypeptide is constitutively active.
4. The method of claim 3, wherein said constitutively active MEF2 polypeptide is a MEF2VP16 fusion protein.
5. The method of claim 3, wherein said constitutively active MEF2 polypeptide contains one or more serinethreonine to aspartic acidglutamic acid substitutions in the MEF2 transactivation domain.
6. The method of claim 1 or claim 3, further comprising inhibiting caspase activity in said progenitor cells.
7. The method of claim 1, wherein said progenitor cells are human stem cells.
8. The method of claim 1, wherein said progenitor cells are embryonic stem cells.
9. The method of claim 8, wherein said embryonic stem cells are human embryonic stem cells.
10. The method of claim 1, wherein said progenitor cells are hematopoietic progenitor cells.
11. The method of claim 10, wherein said hematopoietic progenitor cells are human hematopoietic progenitor cells.
12. The method of claim 1, further comprising selecting CD133-positive human progenitor cells.
13. The method of claim 1, further comprising selecting CD133-positiveCD34-positive human progenitor cells.
14. The method of claim 1, further comprising selecting CD133-positiveCD34-negative human progenitor cells.
15. The method of claim 1, further comprising selecting CD133-positiveCD34-negativeCD45-negative human progenitor cells.
16. The method of claim 1, further comprising selecting CD34-negativeCD38-negativeLin-negative human progenitor cells.
17. The method of claim 1, further comprising selecting CD34-positiveCD38-negativeLin-negativeThy-1-negative human progenitor cells.
18. The method of claim 1, wherein said differentiating agent is retinoic acid.
19. The method of claim 1, wherein said differentiating agent is selected from the group consisting of neurotrophic factor 3, epidermal growth factor, insulin-like growth factor 1 and a platelet-derived growth factor.
20. The method of claim 1, wherein said population containing protected neuronal cells comprises at least 50% neuronal cells.
21. The method of claim 1, further comprising the step of
(c) transplanting cells comprising a nucleic acid molecule encoding a MEF2 polypeptide or an active fragment thereof into a patient to produce a cell population containing protected neuronal cells in said patient.
22. An isolated stem cell, comprising an exogenous nucleic acid molecule encoding a MEF2 polypeptide or an active fragment thereof.
23. The isolated stem cell of claim 22, comprising a nucleic acid molecule encoding a MEF2 polypeptide, or active fragment thereof, operatively linked to a heterologous regulatory element.
24. The isolated stem cell of claim 22, wherein said MEF2 polypeptide is a human MEF2 polypeptide.
25. The isolated stem cell of claim 22, wherein said MEF2 polypeptide is a MEF2C polypeptide.
26. The isolated stem cell of claim 22, wherein said MEF2 polypeptide is constitutively active.
27. The isolated stem cell of claim 26, wherein said MEF2 polypeptide is a constitutively active MEF2C polypeptide.
28. The isolated stem cell of claim 26, wherein said constitutively active MEF2 polypeptide is a MEF2VP16 fusion protein.
29. The isolated stem cell of claim 26, wherein said constitutively active MEF2 polypeptide contains one or more serinethreonine to aspartic acidglutamic acid substitutions in the MEF2 transactivation domain.
30. The isolated stem cell of claim 22, which is a human stem cell.
31. The isolated stem cell of claim 22, which is an embryonic stem cell.
32. The isolated embryonic stem cell of claim 31, which is a human embryonic stem cell.
33. The isolated human stem cell of claim 30, wherein said MEF2 polypeptide is human MEF2C.
34. The isolated human stem cell of claim 30, wherein said MEF2 polypeptide is constitutively active.
35. An isolated hematopoietic stem cell, comprising an exogenous nucleic acid molecule encoding a MEF2 polypeptide or an active fragment thereof.
36. The isolated hematopoietic stem cell of claim 35, comprising a nucleic acid molecule encoding a MEF2 polypeptide, or active fragment thereof, operatively linked to a heterologous regulatory element.
37. The isolated hematopoietic stem cell of claim 36, which is a human hematopoietic stem cell.
38. A method of identifying a protective or differentiation gene, comprising:
(a) isolating a first cell population;
(b) isolating a second cell population, wherein said second cell population has an altered level or activity of a MEF2 polypeptide as compared to said first cell population; and
(c) assaying for differential gene expression in said first cell population as compared to said second cell population,
whereby a gene differentially expressed in said second cell population as compared to said first cell population is identified as a protective or differentiation gene.
39. The method of claim 38, wherein said first cell population is a progenitor cell population, said second cell population is a neuronal cell population, and said differentially expressed gene is a neuronal differentiation gene.
40. The method of claim 38, wherein said first cell population is a progenitor cell population, said second cell population is a muscle cell population, and said differentially expressed gene is a muscle differentiation gene.
41. The method of claim 38, wherein said first and second cell populations are neuronal cell populations, said second cell population has been subject to a neuronal stress as compared to said first cell population, and said differentially expressed gene is a neuroprotective gene.
42. A method of identifying a protective gene in vitro, comprising the steps of:
(a) inducing the p38MEF2 pathway in a cell in vitro to produce a protected cell;
(b) stressing said cell; and
(c) assaying for differential gene expression in said protected cell as compared to gene expression in a control cell,
whereby a gene differentially expressed in said protected cell as compared to said control cell is identified as a protective gene.
43. The method of claim 42, wherein step (a) comprises introducing into said cell a nucleic acid molecule encoding a MEF2 polypeptide.
44. The method of claim 42, wherein said MEF2 polypeptide is a human MEF2 polypeptide.
45. The method of claim 42, wherein said MEF2 polypeptide is a constitutively active MEF2 polypeptide.
46. The method of claim 42, wherein said cell is a neuron.
47. The method of claim 42, wherein said cell is a muscle cell.
48. The method of claim 42, wherein said differential gene expression is increased gene expression.
49. The method of claim 42, wherein said differential gene expression is decreased gene expression.
50. A method of identifying a differentiation gene in vitro, comprising the steps of:
(a) inducing the p38MEF2 pathway in a progenitor cell in vitro to produce a differentiated cell; and
(b) assaying for differential gene expression in said differentiated cell as compared to gene expression in a control cell,
whereby a gene differentially expressed in said differentiated cell as compared to said control cell is identified as a differentiation gene.
51. The method of claim 50, wherein step (a) comprises introducing into said progenitor cell a nucleic acid molecule encoding a MEF2 polypeptide.
52. The method of claim 50, wherein said MEF2 polypeptide is a human MEF2 polypeptide.
53. The method of claim 50, wherein said MEF2 polypeptide is a constitutively active MEF2 polypeptide.
54. The method of claim 50, wherein said differentiated cell is a neuronal cell.
55. The method of claim 50, wherein said differentiated cell is a muscle cell.
56. The method of claim 50, wherein said differential gene expression is increased gene expression.
57. The method of claim 50, wherein said differential gene expression is decreased gene expression.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. In a travel system for an unmanned vehicle that makes the unmanned vehicle travel along a travel route within a work location, a method of controlling travel within the travel system for an unmanned vehicle, comprising the steps of:
generating information on a new travel route by creating topographical data for the work location;
constructing a work location including the new travel route based on the created topographical data;
acquiring actual topographical data for the new travel route by providing the generated information on the new travel route to a vehicle and making the vehicle travel along the new travel route in accordance with temporary travel control data;
correcting the temporary travel control data based on the acquired actual topographical data for the new travel route; and
making the unmanned vehicle travel in accordance with the corrected travel control data.
2. The method of controlling travel within a travel system for an unmanned vehicle according to claim 1, further comprising the step of:
correcting the topographical data for the work location based on the acquired actual topographical data for the new travel route.
3. The method of controlling travel within a travel system for an unmanned vehicle according to claim 1, wherein
topographical data created before construction of the work location includes gradient data, and the temporary travel control data includes a temporary travel speed for the vehicle; and
speed limit by the temporary travel speed is relaxed in accordance with an increase in reliability of the gradient data among the topographical data created before the construction of the work location.
4. The method of controlling travel within a travel system for an unmanned vehicle according to claim 1, wherein
topographical data created before the construction of the work location includes gradient data, and the temporary travel control data includes a temporary travel speed for the vehicle; and
speed limit by the temporary travel speed is relaxed in accordance with an increase in reliability of actual gradient data for the new travel route acquired after the construction of the work location.
5. The method of controlling travel within a travel system for an unmanned vehicle according to claim 1, wherein
topographical data created before the construction of the work location includes gradient data, and the temporary travel control data includes a temporary travel speed for the vehicle;
reliability of the gradient data for the work location is judged by performing acquisition of the actual topographical data a plurality of times; and
speed limit by the temporary travel speed is relaxed in accordance with an increase in reliability of the gradient data for the work location.
6. The method of controlling travel within a travel system for an unmanned vehicle according to claim 1, wherein
both the vehicle for acquiring the actual topographical data for the new travel route and the unmanned vehicle that travels in accordance with the corrected travel control data are unmanned vehicles for carrying freight.
7. A travel system for an unmanned vehicle that makes the unmanned vehicle travel along a travel route within a work location, wherein
the travel system
generates information on a new travel route by creating topographical data for the work location;
constructs the work location including the new travel route based on the created topographical data;
acquires actual topographical data for the new travel route by providing a vehicle with the information on the new travel route generated and making the vehicle travel along the new travel route in accordance with temporary travel control data;
corrects the temporary travel control data based on the acquired actual topographical data for the new travel route; and
makes the unmanned vehicle travel in accordance with the corrected travel control data.
8. The method of controlling travel within a travel system for an unmanned vehicle according to claim 2, wherein
topographical data created before construction of the work location includes gradient data, and the temporary travel control data includes a temporary travel speed for the vehicle; and
speed limit by the temporary travel speed is relaxed in accordance with an increase in reliability of the gradient data among the topographical data created before the construction of the work location.
9. The method of controlling travel within a travel system for an unmanned vehicle according to claim 2, wherein
topographical data created before the construction of the work location includes gradient data, and the temporary travel control data includes a temporary travel speed for the vehicle; and
speed limit by the temporary travel speed is relaxed in accordance with an increase in reliability of actual gradient data for the new travel route acquired after the construction of the work location.
10. The method of controlling travel within a travel system for an unmanned vehicle according to claim 2, wherein
topographical data created before the construction of the work location includes gradient data, and the temporary travel control data includes a temporary travel speed for the vehicle;
reliability of the gradient data for the work location is judged by performing acquisition of the actual topographical data a plurality of times; and
speed limit by the temporary travel speed is relaxed in accordance with an increase in reliability of the gradient data for the work location.
11. The method of controlling travel within a travel system for an unmanned vehicle according to claim 1, wherein
both the vehicle for acquiring the actual topographical data for the new travel route and the unmanned vehicle that travels in accordance with the corrected travel control data are unmanned vehicles for carrying freight.
12. The method of controlling travel within a travel system for an unmanned vehicle according to claim 3, wherein
topographical data created before the construction of the work location includes gradient data, and the temporary travel control data includes a temporary travel speed for the vehicle; and
speed limit by the temporary travel speed is relaxed in accordance with an increase in reliability of actual gradient data for the new travel route acquired after the construction of the work location.
13. The method of controlling travel within a travel system for an unmanned vehicle according to claim 3, wherein
topographical data created before the construction of the work location includes gradient data, and the temporary travel control data includes a temporary travel speed for the vehicle;
reliability of the gradient data for the work location is judged by performing acquisition of the actual topographical data a plurality of times; and
speed limit by the temporary travel speed is relaxed in accordance with an increase in reliability of the gradient data for the work location.
14. The method of controlling travel within a travel system for an unmanned vehicle according to claim 3, wherein
both the vehicle for acquiring the actual topographical data for the new travel route and the unmanned vehicle that travels in accordance with the corrected travel control data are unmanned vehicles for carrying freight.
15. The method of controlling travel within a travel system for an unmanned vehicle according to claim 4, wherein
topographical data created before the construction of the work location includes gradient data, and the temporary travel control data includes a temporary travel speed for the vehicle;
reliability of the gradient data for the work location is judged by performing acquisition of the actual topographical data a plurality of times; and
speed limit by the temporary travel speed is relaxed in accordance with an increase in reliability of the gradient data for the work location.
16. The method of controlling travel within a travel system for an unmanned vehicle according to claim 4, wherein
both the vehicle for acquiring the actual topographical data for the new travel route and the unmanned vehicle that travels in accordance with the corrected travel control data are unmanned vehicles for carrying freight.
17. The method of controlling travel within a travel system for an unmanned vehicle according to claim 5, wherein
both the vehicle for acquiring the actual topographical data for the new travel route and the unmanned vehicle that travels in accordance with the corrected travel control data are unmanned vehicles for carrying freight.
18. The method of controlling travel within a travel system for an unmanned vehicle according to claim 8, wherein
topographical data created before the construction of the work location includes gradient data, and the temporary travel control data includes a temporary travel speed for the vehicle; and
speed limit by the temporary travel speed is relaxed in accordance with an increase in reliability of actual gradient data for the new travel route acquired after the construction of the work location.
19. The method of controlling travel within a travel system for an unmanned vehicle according to claim 8, wherein
topographical data created before the construction of the work location includes gradient data, and the temporary travel control data includes a temporary travel speed for the vehicle;
reliability of the gradient data for the work location is judged by performing acquisition of the actual topographical data a plurality of times; and
speed limit by the temporary travel speed is relaxed in accordance with an increase in reliability of the gradient data for the work location.
20. The method of controlling travel within a travel system for an unmanned vehicle according to claim 8, wherein
both the vehicle for acquiring the actual topographical data for the new travel route and the unmanned vehicle that travels in accordance with the corrected travel control data are unmanned vehicles for carrying freight.