Materials and methods to produce stem cells
First Claim
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1. A method of generating stem cells from a stem cell culture comprising the steps of:
- (i) attaching one or a plurality of stem cells to the surface of a membrane; and
(ii) perfusing the stem cell(s) with culture media.
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Abstract
A method of generating stem cells, such as human and mouse embryonic stem cells, from a stem cell culture comprising the step of perfusing the stem cell culture by flowing culture media through the stem cell culture and apparatus for generating stem cells are disclosed. Also disclosed are methods and apparatus for generating stem cells, such as human and mouse embryonic cells, by culturing the cells on a hydrophilic plastics membrane. Further disclosed is a supplemented conditioned medium for obtaining improved yields of stem cells, such as human and mouse embryonic stem cells.
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Citations
52 Claims
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1. A method of generating stem cells from a stem cell culture comprising the steps of:
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(i) attaching one or a plurality of stem cells to the surface of a membrane; and
(ii) perfusing the stem cell(s) with culture media. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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21. Apparatus for generating stem cells from a stem cell culture by perfusing the stem cells with culture media, the apparatus comprising:
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a membrane configured for attachment of one or more stem cells to be cultured;
culture media supply means for supplying culture media to the stem cell culture;
culture media removal means for removing culture media from said stem cell culture, wherein said supply and removal means are arranged to flow culture media through the stem cell culture. - View Dependent Claims (22, 23)
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24. Apparatus for culturing stem cells comprising:
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(a) a housing having a wall defining an internal chamber;
(b) a stem cell culture membrane extending across said chamber and held in position by;
(b) membrane support means; and
further comprising(c) means to supply culture media to said membrane. - View Dependent Claims (25, 26, 27, 28, 29, 30)
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31. A method of preparing a conditioned medium comprising the steps of:
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(a) culturing support cells in a culture media;
(b) removing a quantity of the culture media; and
(c) diluting the removed culture media. - View Dependent Claims (32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52)
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Specification