Method for monitoring the transition of a cell from one state into another
OwnerKurt Berlin, Alexander Olek +1inventors
InventorsKurt Berlin, Alexander Olek, Sven Olek
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Abstract
The method relates to the field of molecular biology and cell biology. More specifically it is concerned with monitoring a cell's transition from one state into another with the use of a genome based technology. The method is based on a sufficient analysis of methylation patterns according to said cell states. In addition it includes the actual transition of said cell itself. This is done by exposing a cell to conditions expected to convert it from one state to another.
Claims46
A method to monitor the differentiation of at least one cell from a state 1 into a state 2, characterized in that the following steps are carried out
- a) the cytosine methylation pattern of a DNA sample taken from at least one prototype cell at the state 1 is determined or provided,
- b) the cytosine methylation pattern of a DNA sample taken from at least one prototype cell at the state 2 is determined or provided,
- c) at least one cell at the state 1 is exposed to conditions, which are expected to convert a cell at said state 1 into a cell at said state 2,
- d) determining the cytosine methylation pattern in a DNA sample taken from said cell or cells that were exposed to conditions, which are expected to convert a cell at said state 1 into a cell at said state 2,
- e) comparing the cytosine methylation pattern measured in step d) with the cytosine methylation patterns determined or provided in step a) and b) and
- f) concluding whether the conversion of said cell or cells that were exposed to conditions, which are expected to convert a cell at said state 1 into a cell at said state 2 took place, was complete and/or effective.
A method according to the preceding claims, wherein the DNA sample is taken from a source such as a cell culture or a tissue culture.
A method according to the preceding claims, wherein said conditions are characterized as allowing the cells and/or cell cultures to grow on scaffolds or otherwise in 3 dimensional conditions.
A method according to the preceding claims, wherein said conditions include the feeding of said cells on one or several reduced media or supplemented media.
A method according to the preceding claims, wherein said supplemented media contain growth or differentiation inducing factors, natural serums, natural extracts, synthetic supplements, recombinant growth factors or chemicals, which induce growth or differentiation, or a mixture of any of those.
A method according to the preceding claims, wherein said conditions include the feeding of cells on a feeder cell layer.
A method according to the preceding claims, wherein said conditions are characterized by a specified temperature, humidity, light, electrical field, magnetic field, O2, N2 and/or CO2 concentration.
A method according to the preceding claims, wherein said conditions include the treatment of said cells at state 1 with an effective amount of a reagent which is able to modify said cell's DNA methylation status.
A method according to the preceding claims, wherein said conditions include the treatment of said cells at state 1 with an agent involved in DNA methylation belonging to the group of 5-aza-2′-deoxycytidine, Trichostatin A, lankacidin, benzenamine, cyclohexane acetic acid, blue platinum uracil, methyl 13-hydroxy-15-oxo-kaurenoate, sulfonium, euphornin D, octadecylphosphoryl choline, gnidimarcin, and aspiculamycin HCl.
A method according to the preceding claims, wherein said conditions include the treatment of said cells at state 1 with an agent involved in DNA methylation belonging to the group consisting of inhibitors of DNA methylation and histone deacetylation, topoisomerase II, and DNA synthesis.
Dependencies are inferred from the claim text.