Method for isolating and measuring proliferation of long-term label retaining cells and stem cells
OwnerMarc K. Hellerstein, Sylvia Jeewon Kiminventors
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Abstract
This invention relates to a method for separating long-term label retaining cells or stem cells. In particular, this invention relates to a method for separating long-term label retaining cells and/or stem cells from tissues or individuals and for measuring proliferation rates of long-term label retaining cells and stem cells, as well as determining clonal expansion (proliferative history) of cell lineages from the tissues of the individual. The cells may be double-labeled with a cell-lineage marking label and isotopically labeled DNA synthesis precursor prior to physical separation.
Claims45
A method of separating one or more stem cells from a tissue or individual, said method comprising:
- a) administering one or more cell-lineage-marking labels to said tissue or individual for a sufficient time for said one or more cell-lineage marking labels to incorporated into the cells of said tissue or individual;
- b) discontinuing said administering step a) for sufficient time to form a first population of cells and a second population of cells wherein the cells of said second population of cells contain detectably more cell-lineage marking label than the cells of said first population of cells;
- and
- c) detecting the presence or absence of said one or more cell-lineage marking labels in the cells of the first population of cells and the cells of the second population of cells;
- and
- d) separating the cells of said second population of cells, wherein the cells of said second population of cells are stem cells.
A method of determining the proliferation rate of one or more stem cells in a tissue or individual, said method comprising:
- a) administering one or more cell-lineage-marking labels to said tissue or individual for a sufficient time for said one or more cell-lineage marking labels to incorporated in said tissue or individual;
- b) administering one or more isotopically labeled DNA synthesis precursor to said tissue or individual;
- c) discontinuing said administering step a) for sufficient time to form a first population of cells and a second population of cells wherein the cells of said second population of cells contain detectably more cell-lineage marking label than the cells of said first population of cells;
- d) detecting the presence or absence of said one or more cell-lineage marking labels in said the cells of said first population and the cells of said second population;
- e) separating the cells of said second population, wherein the cells of said second population are stem cells;
- and
- f) measuring the isotopic enrichment of one or more deoxyribonucleotides in said stem cells to determine the proliferation rate of one or more stem cells in said tissue or individual.
A kit for separating one or more stem cells in a tissue or individual comprising:
- a) an cell-lineage-marking label, and
- b) instructions for use of the kit,
- wherein said kit is used to separate said one or more stem cells in a tissue or individual.
A kit for determining the proliferation rate of one or more stem cells in a tissue or individual comprising:
- a) an cell-lineage-marking label, and
- b) an isotopically labeled DNA synthesis precursor,
- wherein said kit is used to determine the proliferation rate of one or more stem cells in a tissue or individual.
Dependencies are inferred from the claim text.