Methods and devices for high throughput screening of conditions affecting stem cell differentiation
First Claim
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1. A method for identifying a pluripotency profile of stem cells comprising:
- culturing one or more stem cells in a cell culturing system, wherein the cell culturing system comprises a cell culturing plate comprising a layer of cell culturing substrate, a cell culture site comprising the one or more stem cells, and one or more morphogen sites comprising one or more morphogens, wherein the one or more morphogen sites are each positioned peripheral to the cell culture site, and wherein each of the one or more morphogen sites are separated by physical barriers in the cell culturing plate;
incubating the cell culturing system under conditions suitable to allow the one or more stem cells to differentiate into one or more differentiated cells and migrate on the cell culturing plate in response to the one or more morphogens; and
determining the migration pattern of the one or more differentiated cells on the cell culturing plate, wherein the migration pattern of the one or more differentiated cells is indicative of the pluripotency profile of the population of stem cells.
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Abstract
Disclosed are methods and systems for testing the effects of various morphogens and/or feeder cells on the differentiation of pluripotent cells. The assays described herein can be used for determining the optimum conditions that lead to differentiation of stem cells. Once the optimum conditions for stem cell differentiation are determined, such cells may be used in a variety of therapies.
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Citations
44 Claims
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1. A method for identifying a pluripotency profile of stem cells comprising:
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culturing one or more stem cells in a cell culturing system, wherein the cell culturing system comprises a cell culturing plate comprising a layer of cell culturing substrate, a cell culture site comprising the one or more stem cells, and one or more morphogen sites comprising one or more morphogens, wherein the one or more morphogen sites are each positioned peripheral to the cell culture site, and wherein each of the one or more morphogen sites are separated by physical barriers in the cell culturing plate; incubating the cell culturing system under conditions suitable to allow the one or more stem cells to differentiate into one or more differentiated cells and migrate on the cell culturing plate in response to the one or more morphogens; and determining the migration pattern of the one or more differentiated cells on the cell culturing plate, wherein the migration pattern of the one or more differentiated cells is indicative of the pluripotency profile of the population of stem cells. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A method for identifying a pluripotency profile of stem cells comprising:
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culturing one or more stem cells in a cell culturing system, wherein the cell culturing system comprises a cell culturing plate comprising a layer of cell culturing substrate, a cell culture site comprising the one or more stem cells, and one or more morphogen sites comprising one or more feeder cells, wherein the one or more morphogen sites are each positioned peripheral to the cell culture site; incubating the cell culturing system under conditions suitable for the feeder cells to produce one or more morphogens, and to allow the one or more stem cells to differentiate into one or more differentiated cells and migrate on the cell culturing plate; and determining the migration pattern of the one or more differentiated cells on the cell culturing plate, wherein the migration pattern of the one or more differentiated cells is indicative of the pluripotency profile of the population of stem cells. - View Dependent Claims (14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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26. A method for identifying a feeder a cell population for stem cell differentiation comprising:
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culturing one or more stem cells in a cell culturing system, wherein the cell culturing system comprises a cell culturing plate comprising a layer of cell culturing substrate, a cell culture site comprising the one or more stem cells, and one or more morphogen sites comprising one or more test feeder cells, wherein the one or more morphogen sites are each positioned peripheral to the cell culture site; incubating the cell culturing system under conditions suitable for the feeder cells to produce one or more morphogens, and to allow the one or more stem cells to differentiate into one or more differentiated cells and migrate on the cell culturing plate; and determining the migration pattern of the one or more differentiated cells on the cell culturing plate, wherein the migration pattern of the one or more differentiated cells is indicative of efficacy of test feeder cells on stem cell differentiation. - View Dependent Claims (27, 28)
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29. A method for identifying a stem cell morphogen comprising:
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culturing one or more stem cells in a cell culturing system, wherein the cell culturing system comprises a cell culturing plate comprising a layer of cell culturing substrate, a cell culture site comprising the one or more stem cells, and two or more morphogen sites comprising one or more test agents, wherein each of the two or more morphogen sites is positioned peripheral to the cell culture site, and wherein each of the two or more morphogen sites physically overlap one another; incubating the cell culturing system under conditions suitable to allow the one or more stem cells to differentiate into one or more differentiated cells and migrate on the cell culturing plate in response to the one or more test agents; and determining the migration pattern of the one or more differentiated cells on the cell culturing plate, wherein the migration pattern of the one or more differentiated cells is indicative of test agents'"'"' efficacy as a morphogen. - View Dependent Claims (30, 31)
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32. A cell culturing system comprising a cell culturing plate, wherein the cell culturing plate comprises a layer of cell culturing substrate;
- a cell culture site comprising one or more stem cells; and
two or more morphogen sites, wherein each of the two or more morphogen sites is positioned peripheral to the cell culture site, and wherein each of the two or more morphogen sites physically overlap one another. - View Dependent Claims (33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44)
- a cell culture site comprising one or more stem cells; and
Specification