CN1657604A - Preparation and using method of culture medium - Google Patents
Preparation and using method of culture medium Download PDFInfo
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- CN1657604A CN1657604A CN 200510053211 CN200510053211A CN1657604A CN 1657604 A CN1657604 A CN 1657604A CN 200510053211 CN200510053211 CN 200510053211 CN 200510053211 A CN200510053211 A CN 200510053211A CN 1657604 A CN1657604 A CN 1657604A
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Abstract
A method for preparing and using the culture medium containing glutamine and/or sodium dicarbonate features that the easily decomposed components in the culture medium are individually packed, and then mixed before the culture medium is used. Its advantage is long storage time.
Description
(1) technical field
Invention relates to a kind of culture medium preparation and using method, more specifically to a kind of culture medium preparation and using method that includes glutamine and/or sodium bicarbonate.Belong to field of biological product.
(2) background technology
Substratum is material commonly used in the field of biological product, and is very necessary in bacterium and cell cultures, but certain component of some substratum decomposes easily in the prior art, causes problems such as shelf time weak point, causes the use cost of substratum very high.Wherein, in the animal substratum, glutamine is a kind of component of necessity, but at normal temperatures, and in the very short time, the glutamine composition in the substratum is with regard to degradable more than 90% in day for about 7-10, and this brings very big difficulty to its commercialization.Such substratum is a lot of in the prior art, all has above-mentioned defective, for example, Chinese patent CN1087778C discloses a kind of serum free medium of hybridoma, wherein include the glutamine composition, but the shelf time of this substratum is very limited, is difficult to reach the commercial purpose of using.
(3) summary of the invention
[problem that will solve]
The purpose of this invention is to provide a kind of culture medium preparation and using method that contains easy decomposition composition, especially a kind of culture medium preparation and using method that contains glutamine and/or sodium bicarbonate.
[technical scheme]
Culture medium preparation and using method among the present invention is characterized in that: pack separately easily decomposing component in the liquid nutrient medium, and it is placed under the suitable storage condition separately, before using this substratum two portions are mixed.
Easy decomposition component in the above-mentioned substratum is glutamine and/or sodium bicarbonate;
The storage condition of above-mentioned glutamine and/or sodium bicarbonate concentrated solution be lower than-20 degrees centigrade frozen, preferred-80 degrees centigrade.
The storage condition of the media components of the above-mentioned use that matches with glutamine and/or sodium bicarbonate is 4 to 8 degrees centigrade, preferred 4 degrees centigrade.
[beneficial effect]
The inventive method has good effect, because the material that decomposes just loses its effect after placing for some time easily, can only preserve the very short time, causes unstable product quality.Prepare and use substratum with the inventive method, can prolong the shelf time of substratum, can momentarily use as required, use very conveniently, quality product is more stable, for the commercialization of liquid nutrient medium offers convenience.
(4) embodiment
Following examples are used to illustrate the present invention, but are not used for limiting the scope of the invention.
The present invention is further described with the following examples, but scope of the present invention is not provided constraints.
Embodiment 1: substratum Dulbecco ' s Modified Eagle Medium (D-MEM) (low dextrose)
Be used for animal cell culture, its proportioning is as follows:
| Component | Concentration (mg/L) | Volumetric molar concentration (mM) |
| Glycine (glycine) | ????30 | ????0.400 |
| L-Arginine hydrochloride (L-chlorination arginine) | ????84 | ????0.398 |
| L-Cystine 2HCl (L-2 hydrochloric acid Gelucystine ester) | ????63 | ????0.201 |
| L-Glutamine (L-glutaminate) | ????584 | ????4.00 |
| L-Histidine hydrochloride-H2O (L-one water chlorination Histidine) | ????42 | ????0.200 |
| L-Isoleucine (L-Isoleucine) | ????105 | ????0.802 |
| L-Leucine (L-leucine) | ????105 | ????0.802 |
| L-Lysine hydrochloride (L-chlorination Methionin) | ????146 | ????0.798 |
| L-Methionine (L-methionine(Met)) | ????30 | ????0.201 |
| L-Phenylalanine (L-phenylalanine) | ????66 | ????0.400 |
| L-Serine (L-Serine) | ????42 | ????0.400 |
| L-Threonine (L-Threonine) | ????95 | ????0.798 |
| L-Tryptophan (L-tryptophane) | ????16 | ????0.0784 |
| L-Tyrosine disodium salt dihydrate (L-tyrosine disodium salt) | ????104 | ????0.398 |
| L-Valine (L-Xie Ansuan) | ????94 | ????0.803 |
| Choline chloride (choline chloride 60) | ????4 | ????0.0286 |
| D-Calcium pantothenate (D-calcium pantothenate) | ????4 | ????0.00839 |
| Folic Acid (folic acid) | ????4 | ????0.00907 |
| I-Inositol (i-inositol) | ????7.2 | ????0.0400 |
| Niacinamide (niacinamide) | ????4 | ????0.0328 |
| Pyridoxine hydrochloride (pyridoxine hydrochloride) | ????4 | ????0.0196 |
| Riboflavin (riboflavin) | ????0.4 | ????0.00106 |
| Thiamine hydrochloride (vitamin) | ????4 | ????0.0119 |
| Calcium Chloride (CaCl2) is (Calcium Chloride Powder Anhydrous) (anhyd.) | ????200 | ????1.80 |
| Ferric Nitrate (Fe (NO3) 3 " 9H2O) (9 water iron nitrate) | ????0.1 | ????0.000248 |
| Magnesium Sulfate (MgSO4) is (anhydrous magnesium sulfate) (anhyd.) | ????97.67 | ????0.814 |
| Potassium Chloride (KCl) (Repone K) | ????400 | ????5.33 |
| Sodium Chloride (NaCl) (sodium-chlor) | ????6400 | ????110.34 |
| Sodium Phosphate monobasic (NaH2PO4-H2O) (sodium dihydrogen phosphate-water) | ????125 | ????0.906 |
| D-Glucose (Dextrose) (D-glucose) | ????1000 | ????5.56 |
| Phenol Red (phenol red) | ????15 | ????0.0399 |
| Sodium Pyruvate (Sodium.alpha.-ketopropionate) | ????110 | ????1.000 |
Glutamine in the above-mentioned substratum is packed separately, and preserved down at subzero 80 degrees centigrade.Other components get final product its mixing in use with 4 degrees centigrade of preservations.
Embodiment 2: substratum Dulbecco ' s Modified Eagle Medium (D-MEM) (high glucose), be used for animal cell culture, and its proportioning is as follows:
| Component | Concentration (mg/L) | Volumetric molar concentration (mM) |
| Glycine (glycine) | ????30 | ????0.400 |
| L-Arginine hydrochloride (L-chlorination arginine) | ????84 | ????0.398 |
| L-Cystine 2HCl (L-2 hydrochloric acid Gelucystine ester) | ????63 | ????0.201 |
| L-Glutamine (L-glutaminate) | ????580 | ????3.97 |
| L-Histidine hydrochloride-H2O (L-one water chlorination Histidine) | ????42 | ????0.200 |
| L-Isoleucine (L-Isoleucine) | ????105 | ????0.802 |
| L-Leucine (L-leucine) | ????105 | ????0.802 |
| L-Lysine hydrochloride (L-chlorination Methionin) | ????146 | ????0.798 |
| L-Methionine (L-methionine(Met)) | ????30 | ????0.201 |
| L-Phenylalanine (L-phenylalanine) | ????66 | ????0.400 |
| L-Serine (L-Serine) | ????42 | ????0.400 |
| L-Threonine (L-Threonine) | ????95 | ????0.798 |
| L-Tryptophan (L-tryptophane) | ????16 | ????0.0784 |
| L-Tyrosine (L-tyrosine) | ????72 | ????0.398 |
| L-Valine (L-Xie Ansuan) | ????94 | ????0.803 |
| Choline chloride (choline chloride 60) | ????4 | ????0.0286 |
| D-Calcium pantothenate (D-calcium pantothenate) | ????4 | ????0.00839 |
| Folic Acid (folic acid) | ????4 | ????0.00907 |
| I-Inositol (i-inositol) | ????7.2 | ????0.0400 |
| Niacinamide (niacinamide) | ????4 | ????0.0328 |
| Pyridoxine hydrochloride (pyridoxine hydrochloride) | ????4 | ????0.0196 |
| Riboflavin (riboflavin) | ????0.4 | ????0.00106 |
| Thiamine hydrochloride (vitamin) | ????4 | ????0.0119 |
| Calcium Chloride (CaCl2-2H2O) (Calcium dichloride dihydrate) | ????264 | ????1.80 |
| Ferric Nitrate (Fe (NO3) 3 " 9H2O) (nine water iron nitrates) | ????0.1 | ????0.000248 |
| Magnesium Sulfate (MgSO4-7H2O) (magnesium sulfate heptahydrate) | ????200 | ????0.813 |
| Potassium Chloride (KCl) (Repone K) | ????400 | ????5.33 |
| Sodium Bicarbonate (NaHCO3) (sodium bicarbonate) | ????3700 | ????44.05 |
| Sodium Chloride (NaCl) (sodium-chlor) | ????6400 | ????110.34 |
| Sodium Phosphate monobasic (NaH2PO4-2H2O) (sodium dihydrogen phosphate dihydrate) | ????141 | ????0.916 |
| D-Glucose (Dextrose) (D-glucose) | ????1000 | ????5.56 |
| Phenol Red (phenol red) | ????15 | ????0.0399 |
| Sodium Pyruvate (Sodium.alpha.-ketopropionate) | ????110 | ????1.000 |
Glutamine in the above-mentioned substratum is packed separately, and preserved down at subzero 20 degrees centigrade.Other components get final product its mixing in use with 8 degrees centigrade of preservations.
Embodiment 3 RPMI Medium 1640 are used for animal cell culture, and its proportioning is as follows:
| Component | Concentration (mg/L) | Volumetric molar concentration (mM) |
| Glycine (glycine) | ????10 | ????0.133 |
| L-Arginine (L-arginine) | ????200 | ????1.15 |
| L-Asparagine (L-Radix Asparagi vinegar amino acid) | ????50 | ????0.379 |
| L-Aspartic acid (L-days (door) winter propylhomoserins) | ????20 | ????0.150 |
| L-Cystine 2HCl (L-2 hydrochloric acid Gelucystine ester) | ????65 | ????0.208 |
| L-Glutamic Acid (L-L-glutamic acid) | ????20 | ????0.136 |
| L-Glutamine (L-glutaminate) | ????300 | ??2.05 |
| L-Histidine (L-Histidine) | ????15 | ??0.0968 |
| L-Hydroxyproline (L-hydroxyl (base) proline(Pro)) | ????20 | ??0.153 |
| L-Isoleucine (L-Isoleucine) | ????50 | ??0.382 |
| L-Leucine (L-leucine) | ????50 | ??0.382 |
| L-Lysine hydrochloride (L-chlorination Methionin) | ????40 | ??0.274 |
| L-Methionine (L-methionine(Met)) | ????15 | ??0.101 |
| L-Phenylalanine (L-phenylalanine) | ????15 | ??0.0909 |
| L-Proline (L-proline(Pro)) | ????20 | ??0.174 |
| L-Serine (L-Serine) | ????30 | ??0.286 |
| L-Threonine (L-Threonine) | ????20 | ??0.168 |
| L-Tryptophan (L-tryptophane) | ????5 | ??0.0245 |
| L-Tyrosine disodium salt dihydrate (L-tyrosine disodium salt) | ????29 | ??0.111 |
| L-Valine (L-Xie Ansuan) | ????20 | ??0.171 |
| Biotin (vitamin H) | ????0.2 | ??0.000820 |
| Choline chloride (choline chloride 60) | ????3 | ??0.0214 |
| D-Calcium pantothenate (D-calcium pantothenate) | ????0.25 | ??0.000524 |
| Folic Acid (folic acid) | ????1 | ??0.00227 |
| I-Inositol (i-inositol) | ????35 | ??0.194 |
| Niacinamide (niacinamide) | ????1 | ??0.00820 |
| Para-Aminobenzoic Acid (para-aminobenzoic acid) | ????1 | ??0.00730 |
| Pyridoxine hydrochloride (pyridoxine hydrochloride) | ????1 | ??0.00485 |
| Riboflavin (riboflavin) | ????0.2 | ??0.000532 |
| Thiamine hydrochloride (vitamin) | ????1 | ??0.00297 |
| Vitamin B12 (vitamin B12) | ????0.005 | ??0.0000037 |
| Calcium nitrate (Ca (NO3) 24H2O) (four water-calcium nitrate) | ????100 | ??0.424 |
| Magnesium Sulfate (MgSO4) is (anhydrous magnesium sulfate) (anhyd.) | ????48.84 | ??0.407 |
| Potassium Chloride (KCl) (Repone K) | ????400 | ??5.33 |
| Sodium Bicarbonate (NaHCO3) (sodium bicarbonate) | ????2000 | ??23.81 |
| Sodium Chloride (NaCl) (sodium-chlor) | ????5300 | ??91.38 |
| Sodium Phosphate dibasic (Na2HPO4) anhydrous (AMSP) | ????800 | ??5.63 |
| D-Glucose (Dextrose) (D-glucose) | ????2000 | ???11.11 |
| Glutathione (reduced) (gsh) | ????1 | ???0.00326 |
| ?HEPES | ????5958 | ???25.03 |
| Phenol Red (phenol red) | ????5 | ???0.0133 |
Glutamine in the above-mentioned substratum is packed separately, and preserved down at subzero 50 degrees centigrade.Other components get final product its mixing in use with 6 degrees centigrade of preservations.
Embodiment 4Medium 199 is used for animal cell culture, and its proportioning is as follows:
| Component | Concentration (mg/L) | Volumetric molar concentration (mM) |
| Glycine (glycine) | ????50 | ???0.667 |
| L-Alanine (L-Beta Alanine) | ????25 | ???0.281 |
| L-Arginine hydrochloride (L-chlorination arginine) | ????70 | ???0.332 |
| L-Aspartic acid (L-days (door) winter propylhomoserins) | ????30 | ???0.226 |
| L-Cysteine hydrochloride-H2O (L-one water chlorination halfcystine) | ????0.1 | ???0.000568 |
| L-Cystine 2HCl (L-2 hydrochloric acid Gelucystine ester) | ????26 | ???0.108 |
| L-Glutamic Acid (L-L-glutamic acid) | ????75 | ???0.455 |
| L-Glutamine (L-glutaminate) | ????100 | ???0.685 |
| L-Histidine hydrochloride-H2O (L-one water chlorination Histidine) | ????21.88 | ???0.104 |
| L-Hydroxyproline (L-hydroxyl (base) proline(Pro)) | ????10 | ???0.0763 |
| L-Isoleucine (L-Isoleucine) | ????40 | ???0.305 |
| L-Leucine (L-) leucine) | ????60 | ???0.458 |
| L-Lysine hydrochloride (L-chlorination Methionin) | ????70 | ???0.383 |
| L-Methionine (L-methionine(Met)) | ????15 | ???0.101 |
| L-Phenylalanine (L-phenylalanine) | ????25 | ???0.152 |
| L-Proline (L-proline(Pro)) | ????40 | ???0.348 |
| L-Serine (L-Serine) | ????25 | ???0.238 |
| L-Threonine (L-Threonine) | ????30 | ???0.252 |
| L-Tryptophan (L-tryptophane) | ????10 | ???0.0490 |
| L-Tyrosine disodium salt dihydrate (L-tyrosine disodium salt) | ????40 | ???0.221 |
| L-Valine (L-Xie Ansuan) | ????25 | ???0.214 |
| Alpha-tocopherol Phosphate (alpha-phosphate vitamin-E) | ????0.01 | ???0.0000142 |
| Ascorbic Acid (vitamins C) | ????0.05 | ??0.000284 |
| Biotin (vitamin H) | ????0.01 | ??0.0000410 |
| Choline chloride (choline chloride 60) | ????0.5 | ??0.00357 |
| D-Calcium pantothenate (D-calcium pantothenate) | ????0.01 | ??0.0000210 |
| Folic Acid (vitamins B) | ????0.01 | ??0.0000227 |
| I-Inositol (i-inositol) | ????0.05 | ??0.000278 |
| Menadione (Vitamin K3) (vitamin K3) | ????0.01 | ??0.0000581 |
| Niacinamide (niacinamide) | ????0.025 | ??0.000205 |
| Nicotinic acid (Niacin) (nicotinic acid) | ????0.025 | ??0.000203 |
| Para-Aminobenzoic Acid (para-aminobenzoic acid) | ????0.05 | ??0.000365 |
| Pyridoxal hydrochloride (pyridoxal hydrochloride) | ????0.025 | ??0.000123 |
| Pyridoxine hydrochloride (pyridoxine hydrochloride) | ????0.025 | ??0.000121 |
| Riboflavin (riboflavin) | ????0.01 | ??0.0000266 |
| Thiamine hydrochloride (vitamin) | ????0.01 | ??0.0000297 |
| Vitamin A (acetate) (vitamin A (acetate)) | ????0.1 | ??0.000305 |
| Vitamin D2 (Calciferol) (vitamin D2 (vitamin d)) | ????0.1 | ??0.000252 |
| Calcium Chloride (CaCl2) is (Calcium Chloride Powder Anhydrous) (anhyd.) | ????200 | ??1.80 |
| Ferric nitrate (Fe (NO3)-9H2O) (nine water iron nitrates) | ????0.7 | ??0.00173 |
| Magnesium Sulfate (MgSO4) is (anhydrous magnesium sulfate) (anhyd.) | ????97.67 | ??0.397 |
| Potassium Chloride (KCl) (Repone K) | ????400 | ??5.33 |
| Sodium Bicarbonate (NaHCO3) (sodium bicarbonate) | ????2200 | ??26.19 |
| Sodium Chloride (NaCl) (sodium-chlor) | ????4750 | ??81.90 |
| Sodium Phosphate monobasic (NaH2PO4) anhydrous (AMSP) | ????140 | ??1.01 |
| 2-deoxy-D-ribose (2-deoxy-D-ribose) | ????0.5 | ??0.00373 |
| Adenine sulfate (adenine sulfate) | ????10 | ??0.0248 |
| Adenosine 5 '-phosphate (5 '-adenosine phosphate) | ????0.2 | ??0.000576 |
| Adenosine 5 '-triphosphate (5 '-Triphosaden) | ????1 | ??0.00165 |
| Cholesterol (cholesterol) | ????0.2 | ??0.000517 |
| D-Glucose (Dextrose) (D-glucose) | ????1000 | ??5.56 |
| Glutathione (reduced) (gsh) | ????0.05 | ??0.000163 |
| Guanine hydrochloride (guanine hydrochloride) | ????0.3 | ??0.00160 |
| ?HEPES | ????5960 | ??25.04 |
| Hypoxanthine Na (xanthoglobulin sodium) | ????0.4 | ??0.00294 |
| Phenol Red (phenol red) | ????20 | ??0.0531 |
| Ribose (ribose) | ????0.5 | ??0.00333 |
| Sodium Acetate (sodium acetate) | ????50 | ??0.610 |
| Thymine (thymus pyrimidine) | ????0.3 | ??0.00238 |
| ?Tween?80 | ????20 | |
| Uracil (uridylic) | ????0.3 | ??0.00268 |
| Xanthine-Na (xanthine sodium) | ????0.34 | ??0.00224 |
Glutamine in the above-mentioned substratum is packed separately, and preserved down at subzero 80 degrees centigrade.Other components get final product its mixing in use with 4 degrees centigrade of preservations.
The inventive method and the product that obtains thereof have good effect, prepare and use culture medium with the inventive method, the holding time of energy Extending culture base, can momentarily use as required, use very convenient, reduce use cost, product quality is more stable, for the commercialization of fluid nutrient medium offers convenience.
Claims (10)
1. culture medium preparation and using method is characterized in that: pack separately easily decomposing component in the liquid nutrient medium, and it is placed under the suitable storage condition separately, before using this substratum two portions are mixed.
2. culture medium preparation and using method according to claim 1, it is characterized in that: the easy decomposition component in the substratum wherein is glutamine and/or sodium bicarbonate.
3. as culture medium preparation and using method as described in the claim 2, it is characterized in that: the storage condition of glutamine wherein and/or sodium bicarbonate concentrated solution be lower than-20 degrees centigrade frozen.
4. as culture medium preparation and using method as described in claim 2 or 3, it is characterized in that: the storage condition of glutamine wherein and/or sodium bicarbonate concentrated solution be-80 degrees centigrade frozen.
5. as culture medium preparation and using method as described in the claim 2, it is characterized in that: the storage condition of the media components used of wherein matching with glutamine and/or sodium bicarbonate is 4 to 8 degrees centigrade.
6. as culture medium preparation and using method as described among the claim 1-5 any one, it is characterized in that: wherein the proportioning of substratum is as follows: (mg/L)
Glycine (glycine) 30
L-Arginine hydrochloride (L-chlorination arginine) 84
L-Cystine 2HCl (L-2 hydrochloric acid Gelucystine ester) 63
L-Glutamine (L-glutaminate) 584
L-Histidine hydrochloride-H2O (L-one water chlorination Histidine) 42
L-Isoleucine (L-Isoleucine) 105
L-Leucine (L-leucine) 105
L-Lysine hydrochloride (L-chlorination Methionin) 146
L-Methionine (L-methionine(Met)) 30
L-Phenylalanine (L-phenylalanine) 66
L-Serine (L-Serine) 42
L-Threonine (L-Threonine) 95
L-Tryptophan (L-tryptophane) 16
L-Tyrosine disodium salt dihydrate (L-tyrosine disodium salt) 104
L-Valine (L-Xie Ansuan) 94
Choline chloride (choline chloride 60) 4
D-Calcium pantothenate (D-calcium pantothenate) 4
Folic Acid (folic acid) 4
I-Inositol (i-inositol) 7.2
Niacinamide (niacinamide) 4
Pyridoxine hydrochloride (pyridoxine hydrochloride) 4
Riboflavin (riboflavin) 0.4
Thiamine hydrochloride (vitamin) 4
Calcium Chloride (CaCl2) is (Calcium Chloride Powder Anhydrous) 200 (anhyd.)
Ferric Nitrate (Fe (NO3) 3 " 9H2O) (9 water iron nitrate) 0.1
Magnesium Sulfate (MgSO4) is (anhydrous magnesium sulfate) 97.67 (anhyd.)
Potassium Chloride (KCl) (Repone K) 400
Sodium Chloride (NaCl) (sodium-chlor) 6400
Sodium Phosphate monobasic (NaH2PO4-H2O) (sodium dihydrogen phosphate-water) 125
D-Glucose (Dextrose) (D-glucose) 1000
Phenol Red (phenol red) 15
Sodium Pyruvate (Sodium.alpha.-ketopropionate) 110.
7. as culture medium preparation and using method as described among the claim 1-4 any one, it is characterized in that: wherein the proportioning of substratum is as follows: (mg/L)
Glycine (glycine) 30
L-Arginine hydrochloride (L-chlorination arginine) 84
L-Cystine 2HCl (L-2 hydrochloric acid Gelucystine ester) 63
L-Glutamine (L-glutaminate) 580
L-Histidine hydrochloride-H2O (L-one water chlorination Histidine) 42
L-Isoleucine (L-Isoleucine) 105
L-Leucine (L-shell propylhomoserin) 105
L-Lysine hydrochloride (L-chlorination Methionin) 146
L-Methionine (L-methionine(Met)) 30
L-Phenylalanine (L-phenylalanine) 66
L-Serine (L-Serine) 42
L-Threonine (L-Threonine) 95
L-Tryptophan (L-tryptophane) 16
L-Tyrosine (L-tyrosine) 72
L-Valine (L-Xie Ansuan) 94
Choline chloride (choline chloride 60) 4
D-Calcium pantothenate (D-calcium pantothenate) 4
Folic Acid (folic acid) 4
I-Inositol (i-inositol) 7.2
Niacinamide (niacinamide) 4
Pyridoxine hydrochloride (pyridoxine hydrochloride) 4
Riboflavin (riboflavin) 0.4
Thiamine hydrochloride (vitamin) 4
Calcium Chloride (CaCl2-2H2O) (Calcium dichloride dihydrate) 264
Ferric Nitrate (Fe (NO3) 3 " 9H2O) (nine water iron nitrates) 0.1
Magnesium Sulfate (MgSO4-7H2O) (magnesium sulfate heptahydrate) 200
Potassium Chloride (KCl) (Repone K) 400
Sodium Bicarbonate (NaHCO3) (sodium bicarbonate) 3700
Sodium Chloride (NaCl) (sodium-chlor) 6400
Sodium Phosphate monobasic (NaH2PO4-2H2O) (sodium dihydrogen phosphate dihydrate) 141
D-Glucose (Dextrose) (D-glucose) 1000
Phenol Red (phenol red) 15
Sodium Pyruvate (Sodium.alpha.-ketopropionate) 110.
8. as culture medium preparation and using method as described among the claim 1-4 any one, it is characterized in that: wherein the proportioning of substratum is as follows: (mg/L)
Glycine (glycine) 10
L-Arginine (L-arginine) 200
L-Asparagine (L-Radix Asparagi vinegar amino acid) 50
L-Aspartic acid (L-days (door) winter propylhomoserins) 20
L-Cystine 2HCl (L-2 hydrochloric acid Gelucystine ester) 65
L-Glutamic Acid (L-L-glutamic acid) 20
L-Glutamine (L-glutaminate) 300
L-Histidine (L-Histidine) 15
L-Hydroxyproline (L-hydroxyl (base) proline(Pro)) 20
L-Isoleucine (L-Isoleucine) 50
L-Leucine (L-leucine) 50
L-Lysine hydrochloride (L-chlorination Methionin) 40
L-Methionine (L-methionine(Met)) 15
L-Phenylalanine (L-phenylalanine) 15
L-Proline (L-proline(Pro)) 20
L-Serine (L-Serine) 30
L-Threonine (L-Threonine) 20
L-Tryptophan (L-tryptophane) 5
L-Tyrosine disodium salt dihydrate (L-tyrosine disodium salt) 29
L-Valine (L-Xie Ansuan) 20
Biotin (vitamin H) 0.2
Choline chloride (choline chloride 60) 3
D-Calcium pantothenate (D-calcium pantothenate) 0.25
Folic Acid (folic acid) 1
I-Inositol (i-inositol) 35
Niacinamide (niacinamide) 1
Para-Aminobenzoic Acid (para-aminobenzoic acid) 1
Pyridoxine hydrochloride (pyridoxine hydrochloride) 1
Riboflavin (riboflavin) 0.2
Thiamine hydrochloride (vitamin) 1
Vitamin B12 (vitamin B12) 0.005
Calcium nitrate (Ca (NO3) 24H2O) (four water-calcium nitrate) 100
Magnesium Sulfate (MgSO4) is (anhydrous magnesium sulfate) 48.84 (anhyd.)
Potassium Chloride (KCl) (Repone K) 400
Sodium Bicarbonate (NaHCO3) (sodium bicarbonate) 2000
Sodium Chloride (NaCl) (sodium-chlor) 5300
Sodium Phosphate dibasic (Na2HPO4) anhydrous (AMSP) 800
D-Glucose (Dextrose) (D-glucose) 2000
Glutathione (reduced) (gsh) 1
HEPES?5958
Phenol Red (phenol red) 5.
9. as culture medium preparation and using method as described among the claim 1-4 any one, it is characterized in that: wherein the proportioning of substratum is as follows: (mg/L)
Glycine (glycine) 50
L-Alanine (L-Beta Alanine) 25
L-Arginine hydrochloride (L-chlorination arginine) 70
L-Asparticacid (L-days (door) winter propylhomoserins) 30
L-Cysteine hydrochloride-H2O (L-one water chlorination halfcystine) 0.1
L-Cystine 2HCl (L-2 hydrochloric acid Gelucystine ester) 26
L-Glutamic Acid (L-L-glutamic acid) 75
L-Glutamine (L-glutaminate) 100
L-Histidine hydrochloride-H2O (L-one water chlorination Histidine) 21.88
L-Hydroxyproline (L-hydroxyl (base) proline(Pro)) 10
L-Isoleucine (L-Isoleucine) 40
L-Leucine (L-) leucine) 60
L-Lysine hydrochloride (L-chlorination Methionin) 70
L-Methionine (L-methionine(Met)) 15
L-Phenylalanine (L-phenylalanine) 25
L-Proline (L-proline(Pro)) 40
L-Serine (L-Serine) 25
L-Threonine (L-Threonine) 30
L-Tryptophan (L-tryptophane) 10
L-Tyrosine disodium salt dihydrate (L-tyrosine disodium salt) 40
L-Valine (L-Xie Ansuan) 25
Alpha-tocopherol Phosphate (alpha-phosphate vitamin-E) 0.01
Ascorbic Acid (vitamins C) 0.05
Biotin (vitamin H) 0.01
Choline chloride (choline chloride 60) 0.5
D-Calcium pantothenate (D-calcium pantothenate) 0.01
Folic Acid (vitamins B) 0.01
I-Inositol (i-inositol) 0.05
Menadione (Vitamin K3) (vitamin K3) 0.01
Niacinamide (niacinamide) 0.025
Nicotinic acid (Niacin) (nicotinic acid) 0.025
Para-Aminobenzoic Acid (para-aminobenzoic acid) 0.05
Pyridoxal hydrochloride (pyridoxal hydrochloride) 0.025
Pyridoxine hydrochloride (pyridoxine hydrochloride) 0.025
Riboflavin (riboflavin) 0.01
Thiamine hydrochloride (vitamin) 0.01
Vitamin A (acetate) (vitamin A (acetate)) 0.1
Vitamin D2 (Calciferol) (vitamin D2 (vitamin d)) 0.1
Calcium Chloride (CaCl2) is (Calcium Chloride Powder Anhydrous) 200 (anhyd.)
Ferric nitrate (Fe (NO3)-9H2O) (nine water iron nitrates) 0.7
Magnesium Sulfate (MgSO4) is (anhydrous magnesium sulfate) 97.67 (anhyd.)
Potassium Chloride (KCl) (Repone K) 400
Sodium Bicarbonate (NaHCO3) (sodium bicarbonate) 2200
Sodium Chloride (NaCl) (sodium-chlor) 4750
Sodium Phosphate monobasic (NaH2PO4) anhydrous (AMSP) 140
2-deoxy-D-ribose (2-deoxy-D-ribose) 0.5
Adenine su1fate (adenine sulfate) 10
Adenosine 5 '-phosphate (5 '-adenosine phosphate) 0.2
Adenosine 5 '-triphosphate (5 '-Triphosaden) 1
Cholesterol (cholesterol) 0.2
D-Glucose (Dextrose) (D-glucose) 1000
Glutathione (reduced) (gsh) 0.05
Guanine hydrochloride (guanine hydrochloride) 0.3
HEPES?5960
Hypoxanthine Na (xanthoglobulin sodium) 0.4
Phenol Red (phenol red) 20
Ribose (ribose) 0.5
Sodium Acetate (sodium acetate) 50
Thymine (thymus pyrimidine) 0.3
Tween?8020
Uracil (uridylic) 0.3
Xanthine-Na (xanthine sodium) 0.34.
10. a substratum is characterized in that: with obtaining as any described method among the claim 1-9.
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| CN 200510053211 CN1657604A (en) | 2005-02-07 | 2005-02-07 | Preparation and using method of culture medium |
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| CN 200510053211 CN1657604A (en) | 2005-02-07 | 2005-02-07 | Preparation and using method of culture medium |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107236708A (en) * | 2017-06-23 | 2017-10-10 | 王晓柯 | A kind of serum free medium for supporting HeLa cell attachment cultures |
| EP4036220A1 (en) * | 2015-12-17 | 2022-08-03 | Life Technologies Corporation | Pellets used in cell culture and methods of making thereof |
-
2005
- 2005-02-07 CN CN 200510053211 patent/CN1657604A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4036220A1 (en) * | 2015-12-17 | 2022-08-03 | Life Technologies Corporation | Pellets used in cell culture and methods of making thereof |
| CN107236708A (en) * | 2017-06-23 | 2017-10-10 | 王晓柯 | A kind of serum free medium for supporting HeLa cell attachment cultures |
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