Patent FR-2971850-A1 (FR)
Title: Analyzing multi-stress resistance of animal/vegetable cell by subjecting healthy/cancerous cell or sample with set of non-stressing/stressing effectors, and measuring growth, cellular viability and/or oxidative stress of cell or sample
Inventor: BOULA DE MAREUIL JEAN
IPC: C12Q1/02
Filed: 2011-02-18
Abstract: The method comprises simultaneously or sequentially subjecting a healthy/cancerous cell or sample with a set of non-stressing or stressing effectors, measuring the growth, the cellular viability and/or the oxidative stress of the cell or sample for measuring resistance index of the cells, comparing the results with an identical measurement of the growth or cellular viability of the same type of cell from the patient not provided with the effectors, and applying a thermal-shock with a temperature of 4-75[deg] C. The method comprises simultaneously or sequentially subjecting a healthy/cancerous cell or sample with a set of non-stressing or stressing effectors, measuring the growth, the cellular viability and/or the oxidative stress of the cell or sample for measuring resistance index of the cells, comparing the results with an identical measurement of the growth or cellular viability of the same type of cell from the patient not provided with the effectors, and applying a thermal-shock on the cells with a temperature of 4-75[deg] C. The measurement of the index of resistance includes performing a medical training therapy (MTT) treatment and/or performing the measurement of a metabolism for the generation of reactive oxygen species (ROS). The index resistance corresponds to the measurement of the state of oxidant stress of the cell. The cell is derived from a patient with stressing effectors including prolonged exposure to the sunlight, radiation exposure, contact with carcinogenic agents such as asbestos, smoking, taking drugs, birth control pills, intense practice or inadequately managed in a sport, excessive alcohol consumption, intellectual stress, heat stress, ozone therapy, pollution and infectious agents. The measurement of the growth or cellular viability is carried out by spectroscopic measurements of refraction, diffraction and absorption methods or the quantification of various markers of viability, the growth and cellular death including: MTT test; an expression and/or quantification of specific proteins of a state of the cell including protein53 (p53); the differential expression of enzymes involved in the cellular metabolism and the expression of oxygenated reactive species; the expression of the proteins markers quantified leveled with the DNA or RNA by techniques including PCR or the gene array; and the proteins to be measured by techniques including biochemical, biophysical, immunological or proteomics, where the tests are performed using probes including dihydrorhodamin-123 (DHR123) fluorescent probe for measuring the quantity of produced ROS. The method further comprises withdrawing a sample containing biological material by the application of an adhesive layer on the skin of the patient, removing the adhesive, incubating the adhesive in a solution containing a reactive probe with a diffusible stress in the cells including DHR123, and quantifying the probe react with the solution proportional to the stress using spectrofluorometry method. An independent claim is included for a kit for evaluation of resistance of cells.