Method for the generation of neural progenitor cells
First Claim
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1. A method for obtaining neural progenitor cells, the method comprising the steps of:
- i) dissociation of undifferentiated hBS cells by enzymatic and/or by mechanical treatment to obtain hBS cell aggregates or a mixture of hBS cell aggregates and single cells, ii) seeding the hBS cells from step i) in a growth medium on a support substrate, iii) culturing the hBS cells seeded as in step ii), iv) optionally, replacing a part of the growth medium with a growth medium, v) passaging the obtained cells from step iii) or iv) to obtain neural progenitor cells vi) optionally, seeding the cells from step v) on a support substrate in a growth medium, vii) optionally, repeating step v) and/or vi) at least 1 time.
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Abstract
A rapid, simple and efficient method for the generation of neural progenitor cells from pluripotent/undifferentiated human blastocyst-derived stem (hBS) cells, neural progenitor cells obtained by the method and further differentiation of these cells into the three neural cell lineages, and the use of the neural progenitor cells and the differentiated cells in the preparation of medicaments. An important feature of the method is that neural progenitor cells are produced without a step involving formation of embryoid bodies (EB), improving the efficiency and the reducing the time for generation as compared to known methods.
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74 Claims
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1. A method for obtaining neural progenitor cells, the method comprising the steps of:
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i) dissociation of undifferentiated hBS cells by enzymatic and/or by mechanical treatment to obtain hBS cell aggregates or a mixture of hBS cell aggregates and single cells, ii) seeding the hBS cells from step i) in a growth medium on a support substrate, iii) culturing the hBS cells seeded as in step ii), iv) optionally, replacing a part of the growth medium with a growth medium, v) passaging the obtained cells from step iii) or iv) to obtain neural progenitor cells vi) optionally, seeding the cells from step v) on a support substrate in a growth medium, vii) optionally, repeating step v) and/or vi) at least 1 time. - View Dependent Claims (2, 4, 5, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74)
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3. (canceled)
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6. (canceled)
Specification