Methods for Identifying Stem Cells Based on Nuclear Morphotypes
AssigneeMassachusetts Institute of Technology
InventorsElena V. Gostjeva, William G. Thilly
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Abstract
Methods for identifying stem cells and other cells specific to embryogenesis and carcinogenesis, classifying tissue samples, diagnosing precancerous and cancerous or atherosclerotic lesions, testing the value of anticancer agents, discovering macromolecules specifically expressed in particular cell types, using stem cells in restorative tissue therapy as well as methods for preparing tissue samples so heteromorphic nuclear morphotypes remain intact are disclosed.
Claims37
A method for characterizing the stage of development or pathology of an isolated sample, comprising:
- a) artificially staining the cells and/or macromolecules of the sample, thereby allowing visualization of nuclei, wherein the sample is a cell culture, preneoplastic lesion, tumor sample, or tissue sample;
- b) visualizing the nuclei of cells distributed throughout the sample, wherein the sample is prepared by a method that substantially preserves the integrity of bell-shaped nuclei;
- and
- c) determining the presence and/or absence of a class or classes of nuclear morphotypes including bell-shaped nuclei, wherein the nuclear morphotypes are associated with a stage of development or pathology,
- thereby characterizing the sample based on the presence or absence of a particular class or classes of nuclear morphotypes;
- wherein the stage of development is selected from embryonic, fetal, neonatal, juvenile, or adult stages of development;
- and wherein the pathology is selected from normal, atherosclerotic, preneoplastic, neoplastic and metastatic.
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28-31. (canceled)
A method of identifying one or more anti-tumorigenic agents comprising:
- a) treating a test mammal having a tumor with one or more candidate agents;
- b) determining the nuclear morphology of cells contained within a tumor sample isolated from the test mammal, wherein the cells and/or macromolecules of the sample are artificially stained, thereby allowing visualization of nuclei;
- and
- c) comparing the nuclear morphology of the cells from the test mammal treated with the candidate anti-tumorigenic agent with cells obtained from a control mammal having a tumor but not treated with the candidate anti-tumorigenic agent,
- wherein elimination of cells comprising neoplastic nuclear morphotypes including bell-shaped nuclei in the tumor sample from the test mammal is indicative of the effectiveness of the agent as an anti-tumorigenic agent.
33-37. (canceled)
A method of identifying one or more anti-tumorigenic agents comprising:
- a) treating a sample of cultured tumor cells with one or more candidate agents, wherein the cells and/or macromolecules of the cells are artificially stained, thereby allowing visualization of nuclei;
- and
- b) evaluating the nuclear morphology of the cells contained in the sample, wherein the cells comprise bell-shaped nuclei and in the absence of an anti-tumorigenic agent, the cultured cells maintain their bell-shaped nuclei, and wherein a reduction in the number of bell-shaped nuclei is indicative of the effectiveness of the agent as an anti-tumorigenic agent.
39-41. (canceled)
A method for preparing a mammalian tissue sample suitable for the identification of cells comprising bell-shaped nuclei, comprising:
- a) disrupting cellular adhesions of cellular sheets of the tissue sample;
- b) spreading the cells with disrupted adhesions onto a hard surface;
- and
- c) artificially staining the cells and/or macromolecules of the sample, thereby allowing visualization of nuclei,
- wherein the structural integrity of the nuclei of the cells remains intact, thus rendering the sample suitable for the identification of bell-shaped nuclei.
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46-50. (canceled)
Dependencies are inferred from the claim text.