Culture media for cells originating from the immune system

Claim 1

A medium for culturing cells under serum-free or low serum conditions characterised in that the medium comprises: A. Growth factors selected from the group consisting of: 1. Approximately 0-100 µg/ml insulin 2. Approximately 1-100 µg/ml transferrin 3. Approximately 0-200 µM ethanolamine; B. Approximately 100-1000 µg per ml bovine serum albumin (BSA) complexed in a molar ratio ranging from 1:0.5 to 1:5 with at least one fatty acid selected from the group consisting of: 1. arachadonic acid 2. oleic acid 3. linoleic acid 4. palmitic acid 5. stearic acid 6. linolenic acid 7. palmitoleic acid; C. Amino acids selected from the group consisting of: 1. Approximately 0-100 mg/l L-alanine 2. Approximately 100-1000 mg/1 L-arginine 3. Approximately 0-100 mg/1 L-asparagine 4. Approximately 0-100 mg/l L-aspartic acid 5. Approximately 0-100 mg/1 L-glycine 6. Approximately 7.5-150 mg/1 L-histidine 7. Approximately 10-500 mg/l L-isoleucine 8. Approximately 25-500 mg/l L-leucine 9. Approximately 20-400 mg/l L-lysine HC1 10. Approximately 7.5-150 mg/l L-methionine 11. Approximately 0-150 mg/l L-serine 12. Approximately 10-200 mg/l L-threonine 13. Approximately 7.5-150 mg/l L-phenylalanine 14. Approximately 2.5-25 mg/l L-tryptophan 15. Approximately 10-100 mg/l L-valine 16. Approximately 0-100 mg/l L-glutamate 17. Approximately 0-200 mg/l L-proline 18. Approximately 0-100 mg/l hydroxy-L-proline 19. Approximately 10-100 mg/l tyrosine 20. Approximately 25-500 mg/l cystine 21. Approximately 100-3000 mg/l glutamine 22. Approximately 0-5.5 mg/l L-α-amino-n-butyric acid 23. Approximately 0-3.9 mg/l D-glucosamine HCl 24. Approximately 0-4.2 mg/l L-taurine 25. Approximately 0-9.4 mg/l L-ornithine HC1; D.Vitamins selected from the group consisting of: 1. Approximately 0-5 mg/l p-amino benzoic acid 2. Approximately 0-3 mg/l biotin 3. Approximately 0.1-3 mg/l d- C a-pantothenate 4. Approximately 1.0-10.0 mg/l choline chloride 5. Approximately 0.5-5 mg/l nicotinamide 6. Approximately 0.5-3 mg/l pyridoxine-HCl 7. Approximately 0.1-1.0 mg/1 riboflavin 8. Approximately 0.5-3.0 mg/l thiamine HC1 9. Approximately 0-10.0 mg/l vitamin B 12 10. Approximately 10.0-50.0 mg/l inositol 11. Approximately 0-0.025 mg/1 D-tacopherol acid succinate 12. Approximately 0-0.25 mg/1 calciferol 13. Approximately 0-0.025 mg/l menadione 14. Approximately 0-0.0625 mg/l niacin 15. Approximately 0-0.25 mg/l vitamin A 16. Approximately 0.5-3.0 mg/l folic acid; E. A balanced salt solution; F. Trace compounds selected from the group consisting of: 1. Approximately 0-0.01 mg/l CuS0 4 .5H 2 0 2. Approximately 0-1x10 -8 M FeS0 4 .7H 2 0 3. Approximately 0-0.1 mg/l ZnS0 4 .7H 2 0 4. Approximately 0-1x10 -7 M L iCl 5. Approximately 0-2.5x10 -6 M Na 2 SeO 4 ; G. Coenzymes selected from the group consisting of: 1. Approximately 0-0.25 mg/l coenzyme A 2. Approximately 0-0.7 mg/l diphosphopyridine nucleotide 3. Approximately 0-1.0 mg/l flavin adenine dinucleotide 4. Approximately 0-1.0 mg/l triphosphopyridine nucleotide 5. Approximately 0-1.0 mg/l cocarboxylase 6. Approximately 0-1.0 mg/l uridine triphosphate, sodium; H. Nucleic acid derivatives selected from the group consisting of: 1. Approximately 0-10 mg/l deoxyadenosine 2. Approximately 0-10 mg/l deoxyguanosine 3. Approximately 0-10 mg/l deoxycytidine HC1 4. Approximately 0-0.1 mg/1 5-methyl cytosine 5. Approximately 0-73 mg/l thymidine; and I. Additional components selected from the group consisting of: 1. Approximately 0-20 mg/l lipoic acid 2. Approximately 0-1000 mg/l sodium pyruvate 3. Approximately 0-40 mg/l hypoxanthine 4. Approximately 2000 mg/l NaHCO 3 5. Approximately 1000-4500 mg/l glucose 6. Approximately 0-20 mg/l phenol red 7. Approximately 0-10 mg/l gluthathione 8. Approximately 0.50 mg/l ascorbic acid 9. Approximately 0-1.8 mg/l D-glucuronolactone 10. Approximately 0-1.8 mg/l sodium glucuronate H 2 0 11. Approximately 0-12.5 mg/l Tween 80.

Claim 2

A medium for culturing cells under serum-free or low serum conditions characterised in that the medium comprises: A. Growth factors selected from the group consisting of: 1. Approximately 0-100 µg/ml insulin 2. Approximately 1-100 µg/ml transferrin 3. Approximately 0-200 µM ethanolamine; B. Approximately 100-1000 µg per ml bovine serum albumin (BSA) complexed in a molar ratio ranging from 1:0.5 to 1:5 with at least one fatty acid selected from the group consisting of: 1. arachadonic acid 2. oleic acid 3. linoleic acid 4. palmitic acid 5. stearic acid 6. linolenic acid 7. palmitoleic acid; C. Amino acids selected from the group consisting of: 1. Approximately 0-100 mg/l L-alanine 2. Approximately 100-1000 mg/1 L-arginine 3. Approximately 0-100 mg/1 L-asparagine 4. Approximately 0-100 mg/l L-aspartic acid 5. Approximately 0-100 mg/1 L-glycine 6. Approximately 7.5-150 mg/1 L-histidine 7. Approximately 10-500 mg/l L-isoleucine 8. Approximately 25-500 mg/l L-leucine 9. Approximately 20-400 mg/l L-lysine HC1 10. Approximately 7.5-150 mg/l L-methionine 11. Approximately 0-150 mg/l L-serine 12. Approximately 10-200 mg/l L-threonine 13. Approximately 7.5-150 mg/l L-phenylalanine 14. Approximately 2.5-25 mg/l L-tryptophan 15. Approximately 10-100 mg/l L-valine 16. Approximately 0-100 mg/l L-glutamate 17. Approximately 0-200 mg/l L-proline 18. Approximately 0-100 mg/l hydroxy-L-proline 19. Approximately 10-100 mg/l tyrosine 20. Approximately 25-500 mg/l cystine 21. Approximately 100-3000 mg/l glutamine 22. Approximately 0-5.5 mg/l L-α-amino-n-butyric acid 23. Approximately 0-3.9 mg/l D-glucosamine HCl 24. Approximately 0-4.2 mg/l L-taurine 25. Approximately 0-9.4 mg/l L-ornithine HC1; D.Vitamins selected from the group consisting of: 1. Approximately 0-5 mg/l p-amino benzoic acid 2. Approximately 0-3 mg/l biotin 3. Approximately 0.1-3 mg/l d- C a-pantothenate 4. Approximately 1.0-10.0 mg/l choline chloride 5. Approximately 0.5-5 mg/l nicotinamide 6. Approximately 0.5-3 mg/l pyridoxine-HCl 7. Approximately 0.1-1.0 mg/1 riboflavin 8. Approximately 0.5-3.0 mg/l thiamine HC1 9. Approximately 0-10.0 mg/l vitamin B 12 10. Approximately 10.0-50.0 mg/l inositol 11. Approximately 0-0.025 mg/1 D-tacopherol acid succinate 12. Approximately 0-0.25 mg/1 calciferol 13. Approximately 0-0.025 mg/l menadione 14. Approximately 0-0.0625 mg/l niacin 15. Approximately 0-0.25 mg/l vitamin A 16. Approximately 0.5-3.0 mg/l folic acid; E. A balanced salt solution; F. Trace compounds selected from the group consisting of: 1. Approximately 0-0.01 mg/l CuS0 4 .5H 2 0 2. Approximately 0-1x10 -8 M FeS0 4 .7H 2 0 3. Approximately 0-0.1 mg/l ZnS0 4 .7H 2 0 4. Approximately 0-1x10 -7 M L iCl 5. Approximately 0-2.5x10 -6 M Na 2 SeO 4 ; G. Coenzymes selected from the group consisting of: 1. Approximately 0-0.25 mg/l coenzyme A 2. Approximately 0-0.7 mg/l diphosphopyridine nucleotide 3. Approximately 0-1.0 mg/l flavin adenine dinucleotide 4. Approximately 0-1.0 mg/l triphosphopyridine nucleotide 5. Approximately 0-1.0 mg/l cocarboxylase 6. Approximately 0-1.0 mg/l uridine triphosphate, sodium; H. Nucleic acid derivatives selected from the group consisting of: 1. Approximately 0-10 mg/l deoxyadenosine 2. Approximately 0-10 mg/l deoxyguanosine 3. Approximately 0-10 mg/l deoxycytidine HC1 4. Approximately 0-0.1 mg/1 5-methyl cytosine 5. Approximately 0-73 mg/l thymidine; and I. Additional components selected from the group consisting of: 1. Approximately 0-20 mg/l lipoic acid 2. Approximately 0-1000 mg/l sodium pyruvate 3. Approximately 0-40 mg/l hypoxanthine 4. Approximately 2000 mg/l NaHCO 3 5. Approximately 1000-4500 mg/l glucose 6. Approximately 0-20 mg/l phenol red 7. Approximately 0-10 mg/l gluthathione 8. Approximately 0.50 mg/l ascorbic acid 9. Approximately 0-1.8 mg/l D-glucuronolactone 10. Approximately 0-1.8 mg/l sodium glucuronate H 2 0 11. Approximately 0-12.5 mg/l Tween 80.

Claim 3

A medium culturing cells under serum-free or low serum conditions characterised in that the medium comprises: A. Growth factors selected from the group consisting of: 1. Approximately 5 µg/ml insulin 2. Approximately 50 µg/ml transferrin 3. Approximately 20 µM ethanolamine; B. Approximately 100-1000 µg/ml of Bovine serum albumin (BSA) complexed in a 1:1 molar ratio with each of the following fatty acids: 1. arachadonic acid 2. oleic acid 3. linoleic acid 4. palmitic acid 5. stearic acid 6. linolenic acid 7. palmitoleic acid; C. Amino acids selected from the group consisting of: 1. Approximately 10 mg/l L-alanine 2. Approximately 200 mg/l L-arginine 3. Approximately 50 mg/l L-asparagine 4. Approximately 20 mg/l L-aspartic acid 5. Approximately 10 mg/1 L-glycine 6. Approximately 15 mg/l L-histidine 7. Approximately 50 mg/l L-isoleucine 8. Approximately 50 mg/l L-leucine 9. Approximately 40 mg/l L-lysine HC1 10. Approximately 15 mg/l L-methionine 11. Approximately 30 mg/l L-serine 12. Approximately 20 mg/l L-threonine 13. Approximately 15 mg/l L-phenylalanine 14. Approximately 5 mg/l L-tryptophan 15. Approximately 20 mg/l L-valine 16. Approximately 20 mg/l L-glutamate 17. Approximately 20 mg/l L-proline 18. Approximately 20 mg/l hydroxy-L-proline 19. Approximately 20 mg/l tyrosine 20. Approximately 50 mg/l cystine 21. Approximately 300 mg/l glutamine 22. Approximately 0.55 mg/l L-α-amino-n-butyric acid 23. Approximately 0.39 mg/l D-glucosamine HCl 24. Approximately 0.42 mg/l L-taurine 25. Approximately 0.94 mg/l L-ornithine HC1; D. Vitamins selected from the group consisting of: 1. Approximately 1 mg/l p-aminobenzoic acid 2. Approximately 0.2 mg/l biotin 3. Approximately 0.25 mg/l d-Ca-pantothenate 4. Approximately 3 mg/l choline chloride 5. Approximately 1 mg/l nicotinamide 6. Approximately 1 mg/l pxridoxine-HCl 7. Approximately 0.2 mg/l riboflavin 8. Approximately 1 mg/l thiamine HC1 9 . Approximately 1.0 mg/l vitamin B 12 10. Approximately 35 mg/l inositol 11. Approximately 0.0025 mg/l D-tacopherol acid succinate 12. Approximately 0.025 mg/l calciferol 13. Approximately 0.0025 mg/l menadione 14. Approximately 0.00625 mg/l niacin 15. Approximately 0.025 mg/l vitamin A 16. Approximately 1 mg/l folic acid; E. A balanced salt solution; F. Trace compounds selected from the group consisting of: 1. Approximately 0.00125 mg/l CuS0 4 .5H 2 0 2. Approximately 10 -10 M FeSO 4 ·7H 2 O 3. Approximately 0.028 mg/l ZnS0 4 .7H 2 0 4. Approximately 10 -9 M LiCl 5. Approximately 2.5x10 -8 M Na 2 SeO 4 ; G. Coenzymes selected from the group consisting of: 1. Approximately 0.025 mg/l coenzyme A 2. Approximately 0.07 mg/l diphosphorpyridine nucleotide 3. Approximately 0.1 mg/l flavin adenine dinucleotide 4. Approximately 0.1 mg/l triphosphopyridine nucleotide 5. Approximately 0.1 mg/l cocarboxylase 6. Approximately 0.1 mg/l uridine triphosphate sodium; H . Nucleic acid derivatives selected from the group consisting of: 1. Approximately 1 mg/l deoxyadenosine 2. Approximately 1 mg/l deoxyguanosine 3. Approximately 1 mg/l deoxycytidine HC1 4. Approximately 0.02 mg/1 5-methyl cytosine 5. Approximately 1 mg/l thymidine; and I. Additional components selected from the group consisting of: 1. Approximately 0.2 mg/l lipoic acid 2. Approximately 110 mg/l sodium pyruvate 3. Approximately 4 mg/l hypoxanthine 4. Approximately 200 mg/l NaHCO 3 5. Approximately 2000 mg/l glucose 6. Approximately 5 mg/l phenol red 7. Approximately 1 mg/l gluthathione 8. Approximately 5 mg/l ascorbic acid 9. Approximately 0.18 mg/l D-glucuronolactone 10. Approximately 0.18 mg/l sodium glucuronate H 2 0 11. Approximately 1.25 mg/l Tween 80.

Claim 4

A medium culturing cells under serum-free or low serum conditions characterised in that the medium comprises: A. Growth factors selected from the group consisting of: 1. Approximately 5 µg/ml insulin 2. Approximately 50 µg/ml transferrin 3. Approximately 20 µM ethanolamine; B. Approximately 100-1000 µg/ml of Bovine serum albumin (BSA) complexed in a 1:1 molar ratio with each of the following fatty acids: 1. arachadonic acid 2. oleic acid 3. linoleic acid 4. palmitic acid 5. stearic acid 6. linolenic acid 7. palmitoleic acid; C. Amino acids selected from the group consisting of: 1. Approximately 10 mg/l L-alanine 2. Approximately 200 mg/l L-arginine 3. Approximately 50 mg/l L-asparagine 4. Approximately 20 mg/l L-aspartic acid 5. Approximately 10 mg/1 L-glycine 6. Approximately 15 mg/l L-histidine 7. Approximately 50 mg/l L-isoleucine 8. Approximately 50 mg/l L-leucine 9. Approximately 40 mg/l L-lysine HC1 10. Approximately 15 mg/l L-methionine 11. Approximately 30 mg/l L-serine 12. Approximately 20 mg/l L-threonine 13. Approximately 15 mg/l L-phenylalanine 14. Approximately 5 mg/l L-tryptophan 15. Approximately 20 mg/l L-valine 16. Approximately 20 mg/l L-glutamate 17. Approximately 20 mg/l L-proline 18. Approximately 20 mg/l hydroxy-L-proline 19. Approximately 20 mg/l tyrosine 20. Approximately 50 mg/l cystine 21. Approximately 300 mg/l glutamine 22. Approximately 0.55 mg/l L-α-amino-n-butyric acid 23. Approximately 0.39 mg/l D-glucosamine HCl 24. Approximately 0.42 mg/l L-taurine 25. Approximately 0.94 mg/l L-ornithine HC1; D. Vitamins selected from the group consisting of: 1. Approximately 1 mg/l p-aminobenzoic acid 2. Approximately 0.2 mg/l biotin 3. Approximately 0.25 mg/l d-Ca-pantothenate 4. Approximately 3 mg/l choline chloride 5. Approximately 1 mg/l nicotinamide 6. Approximately 1 mg/l pxridoxine-HCl 7. Approximately 0.2 mg/l riboflavin 8. Approximately 1 mg/l thiamine HC1 9 . Approximately 1.0 mg/l vitamin B 12 10. Approximately 35 mg/l inositol 11. Approximately 0.0025 mg/l D-tacopherol acid succinate 12. Approximately 0.025 mg/l calciferol 13. Approximately 0.0025 mg/l menadione 14. Approximately 0.00625 mg/l niacin 15. Approximately 0.025 mg/l vitamin A 16. Approximately 1 mg/l folic acid; E. A balanced salt solution; F. Trace compounds selected from the group consisting of: 1. Approximately 0.00125 mg/l CuS0 4 .5H 2 0 2. Approximately 10 -10 M FeSO 4 ·7H 2 O 3. Approximately 0.028 mg/l ZnS0 4 .7H 2 0 4. Approximately 10 -9 M LiCl 5. Approximately 2.5x10 -8 M Na 2 SeO 4 ; G. Coenzymes selected from the group consisting of: 1. Approximately 0.025 mg/l coenzyme A 2. Approximately 0.07 mg/l diphosphorpyridine nucleotide 3. Approximately 0.1 mg/l flavin adenine dinucleotide 4. Approximately 0.1 mg/l triphosphopyridine nucleotide 5. Approximately 0.1 mg/l cocarboxylase 6. Approximately 0.1 mg/l uridine triphosphate sodium; H . Nucleic acid derivatives selected from the group consisting of: 1. Approximately 1 mg/l deoxyadenosine 2. Approximately 1 mg/l deoxyguanosine 3. Approximately 1 mg/l deoxycytidine HC1 4. Approximately 0.02 mg/1 5-methyl cytosine 5. Approximately 1 mg/l thymidine; and I. Additional components selected from the group consisting of: 1. Approximately 0.2 mg/l lipoic acid 2. Approximately 110 mg/l sodium pyruvate 3. Approximately 4 mg/l hypoxanthine 4. Approximately 200 mg/l NaHCO 3 5. Approximately 2000 mg/l glucose 6. Approximately 5 mg/l phenol red 7. Approximately 1 mg/l gluthathione 8. Approximately 5 mg/l ascorbic acid 9. Approximately 0.18 mg/l D-glucuronolactone 10. Approximately 0.18 mg/l sodium glucuronate H 2 0 11. Approximately 1.25 mg/l Tween 80.