Stem cell culture medium and method of using said medium and the cells
First Claim
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1. A method of enriching stem cells in a plurality of cells, comprising the steps of:
- adding a sample of cells to a first liquid cell culture medium under conditions wherein stem cells are substantially in suspension and wherein non-proliferating cells substantially adhere to a substrate; and
passaging suspended cells from the first medium into a second liquid cell culture medium, thereby enriching the stem cells.
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Abstract
The present invention relates to methods and compositions concerning isolation of proliferating cells. In particular, the invention regards enrichment of stem cells in a mixture of stem cells and non-stem cells, wherein the non-stem cells may be differentiated cells. The invention exploits the non-adherent property of stem cells, as opposed to the adherent property of differentiating cells, by serially passaging the suspended cells in liquid media.
31 Citations
33 Claims
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1. A method of enriching stem cells in a plurality of cells, comprising the steps of:
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adding a sample of cells to a first liquid cell culture medium under conditions wherein stem cells are substantially in suspension and wherein non-proliferating cells substantially adhere to a substrate; and
passaging suspended cells from the first medium into a second liquid cell culture medium, thereby enriching the stem cells. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method of treating an individual for a medical condition, comprising the steps of:
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adding a stem cell-comprising sample of cells to a first liquid cell culture medium under conditions wherein the stem cells are substantially in suspension and wherein non-proliferating cells substantially adhere to a substrate;
passaging the suspended cells from the first medium into a second liquid cell culture medium; and
delivering one or more stem cells to the individual. - View Dependent Claims (16, 17, 18, 19, 20, 21, 22, 23)
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24. A method of isolating one or more mammalian stem cells, comprising the steps of:
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providing a plurality of cells, said plurality comprising one or more stem cells;
subjecting the plurality of cells to a culturing step in a container comprising a liquid cell medium, said culturing producing suspended cells and container-adherent cells;
transferring a plurality of suspended cells to another container comprising a liquid cell medium; and
repeating the subjecting and tranferring steps at least once. - View Dependent Claims (25, 26, 27, 28, 29, 30)
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31. One or more isolated stem cells produced by the steps of:
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adding a sample of cells to a first liquid cell culture medium under conditions wherein stem cells are substantially in suspension in the first medium and wherein non-proliferating cells substantially adhere to a substrate; and
passaging suspended cells into a second liquid cell culture medium.
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32. A method of treating an individual for a medical condition, comprising the steps of:
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adding a stem cell-comprising sample of cells to a first liquid cell culture medium under conditions wherein the stem cells are substantially in suspension and wherein non-proliferating cells substantially adhere to a substrate;
passaging the suspended cells, said suspended cells comprising one or more stem cells, from the first medium into at least a second liquid cell culture medium;
delivering one or more therapeutic agents to one or more of the stem cells, wherein the therapeutic agent is suitable for the condition; and
delivering one or more therapeutic agent-comprising stem cells to the individual. - View Dependent Claims (33)
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Specification