# US 2014/0154672 A1 · Methods for Identifying Stem Cells Based on Nuclear Morphotypes > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2014/0154672 A1 - Document ID: US20140154672A1 - Status: Published application - Kind: A1 - Application number: 13/914,152 - Filed: 2013-06-10 - Published: 2014-06-05 - Earliest claimed: 2004-06-17 - Assignees: Massachusetts Institute of Technology - Applicants: Massachusetts Institute of Technology - Inventors: Elena V. Gostjeva; William G. Thilly - Web page: https://patentgrep.com/patent/US20140154672A1 ## CPC - C12Q 1/6881: for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes - G01N 1/312: {for samples mounted on planar substrates} - G01N 33/5011: {for testing antineoplastic activity} - G01N 33/5017: {for testing neoplastic activity} - G01N 33/5026: {on cell morphology} - G01N 33/5091: {for testing the pathological state of an organism} - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## 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. ## Claims ### Claim 1 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. ### Claim 2 (depends on 1) The method of claim 1, wherein the sample is a tissue sample obtained by surgical excision. ### Claim 3 (depends on 1) The method of claim 1, wherein the sample is physically or chemically fixed prior to cellular degradation of nuclei. ### Claim 4 (depends on 3) The method of claim 3, wherein the sample is frozen. ### Claim 5 (depends on 3) The method of claim 3, wherein the sample is treated with one or more chemical fixing agents selected from the group consisting of: alcohols, aldehydes, organic acids and combinations thereof. ### Claim 6 (depends on 5) The method of claim 5, wherein the fixing agent comprises methanol and acetic acid. ### Claim 7 (canceled) ### Claim 8 (depends on 1) The method of claim 1, wherein the cells of the sample are partially dissociated by tissue maceration and spreading. ### Claim 9 (canceled) ### Claim 10 (depends on 1) The method of claim 1, wherein DNA is stained, thereby allowing visualization of nuclei. ### Claim 11 (depends on 2) The method of claim 2, wherein the tissue sample is fixed within 30 minutes of surgical removal. ### Claim 12 (depends on 1) The method of claim 1, wherein the tissue sample is obtained from a mammal. ### Claim 13 (canceled) ### Claim 14 (canceled) ### Claim 15 (depends on 12) The method of claim 12, wherein the mammal is selected from the group consisting of: primates, rodents, canines, felines, porcines, ovines, bovines and rabbits. ### Claim 16 (depends on 15) The method of claim 15, wherein the mammal is a human. ### Claim 17 (canceled) ### Claim 18 (canceled) ### Claim 19 (depends on 1) The method of claim 1, wherein the class or classes of nuclear morphotypes further comprise cigar-shaped, condensed spherical, spherical, oval, sausage-shaped, kidney-shaped, and bullet-shaped nuclei. ### Claim 20 (depends on 1) The method of claim 1, further comprising determining the spatial or numerical distribution of one or more classes of nuclear morphotypes within the sample, wherein the spatial or numerical distributions of the one or more classes of nuclear morphotypes further characterizes the sample. ### Claim 21 (canceled) ### Claim 22 (depends on 1) The method of claim 1, wherein the nuclei are contained in multinuclear syncytia or in mononuclear cells. ### Claim 23 (canceled) ### Claim 24 (canceled) ### Claim 25 (depends on 1) The method of claim 1, wherein structures indicative of amitotic symmetrical nuclear division are indicative of neoplasia or metastasis. ### Claim 26 (depends on 22) The method of claim 22, wherein the presence of bell-shaped nuclei, the absence of amitotic symmetrical nuclear division and the absence of multinuclear syncytia is indicative of preneoplasia. ### Claim 27 (depends on 1) The method of claim 1, wherein the presence of non-spherical and non-oval nuclei in blood vessel wall tissue is indicative of an incipient atherosclerotic condition. ### Claim at position 28 28-31. (canceled) ### Claim 32 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. ### Claim at position 30 33-37. (canceled) ### Claim 38 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. ### Claim at position 32 39-41. (canceled) ### Claim 42 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. ### Claim 43 (canceled) ### Claim 44 (depends on 42) The method of claim 42, wherein the tissue sample forms a layer on the microscope slide of about 0.5 millimeters. ### Claim 45 (depends on 42) The method of claim 42, wherein the tissue sample forms a layer greater than about 50 microns. ### Claim at position 37 46-50. (canceled)