Patent ID: 7393685

Claim:
A method of cultivating cancer cells from human tissue for molecular-biological mass screenings wherein a tissue sample is locally separated into disk segments by sequential and parallel mechanical splitting based on its heterogeneous structure of tumor cells, normal cells, and contaminants, and wherein said separated tissue sample segments are further split into tissue fragments, and wherein said separated tissue fragments and fluids of the locally separated tissue sample segments are selectively cultivated in a specific culture medium for cultivating tumor cells and under predefined cultivation conditions and under suppression of the disturbing influence or normal cells and contaminants, and wherein the tissue fragments and fluids obtained from the locally separated tissue sample segments are cultivated separately in cell culture bottles filled with said culture medium for cultivating tumor cells and coated with a biomatrix substrate in an 0.01% to 3% oxygen atmosphere, an 0.1% to 5% carbon dioxide atmosphere, at a humidity of 100% and temperatures in the range from 30° C. to 36.5° C. and wherein said tissue sample is temporarily placed in a culture medium for storage together with adhering erythrocytes from the collection site of the sample in the respective patient until the tissue sample fragments are produced, and wherein the tissue sample is kept in the culture medium for storage for a minimum of 2 hours but no longer than 24 hours at temperature in the range from 4° C. to 12° C. to get adapted to said culture medium for storage, and wherein the culture medium for cultivating tumors is composed of inorganic salts, specifically Ca(NO 3 ) 2 50 mg/L CaCl 2 •2H 2 O 132 mg/L KCl 400 mg/L MgSO 4 •7H 2 O 150 mg/L NaCl 6400 mg/L NaHCO 3 2100 mg/L Na 2 HPO 4 400 mg/L; amino acids, specifically L-arginine•4HCl 110 mg/L L-asparagine (free base) 38 mg/L L-aspartic acid 23 mg/L L-cystine 31 mg/L L-glutamic acid 25 mg/L L-glutamine 296 mg/L Glycine 13 mg/L L-histidine (free base) 12 mg/L L-hydroxyproline 10 mg/L L-isoleucine 38 mg/L L-leucine 38 mg/L L-lysine HCl 35 mg/L L-methionine 12 mg/L L-phenylalanine 16 mg/L L-proline 22 mg/L L-serine 26 mg/L L-threonine 22 mg/L L-tryptophane 5 mg/L L-tyrosine 19 mg/L L-valine 22 mg/L L-alanine 10 mg/L vitamins, specifically Biotin 0.6 mg/L D-Ca-pantothenate 0.7 mg/L Choline chloride 3.5 mg/L Folic acid 1.0 mg/L i-Inositol 35.9 mg/L Niacin amide 1.0 mg/L Pyridoxine•HCl 1.0 mg/L Riboflavin 20 mg/L Thiamine•HCl 1.0 mg/L Paraminobenzoic acid 500 mg/L Vitamin B 12 5 mg/L Niacin 25 mg/L Ascorbic acid 50 mg/L Folinic acid 6 mg/L Liponic acid 21 mg/L Vitamin A (acetate) 100 mg/L Pyridoxine•HCl 25 mg/L Niacinamide 25 mg/L a-Tocopherol phosphate 10 mg/L and D-glucose 1750 mg/L Phenol red 7 mg/L Glutathione (reduced) 0.5 mg/L Sodium pyruvate 1 nM Epidermal growth factor (EGF) 250 ng/L Fetal bovine serum (FBS) 12.5% 8 mg/L Bovine insulin (lyophilisate and antibiotics.