CN107217030A - A kind of wide spectrum cell culture medium - Google Patents

A kind of wide spectrum cell culture medium Download PDF

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CN107217030A
CN107217030A CN201710525237.5A CN201710525237A CN107217030A CN 107217030 A CN107217030 A CN 107217030A CN 201710525237 A CN201710525237 A CN 201710525237A CN 107217030 A CN107217030 A CN 107217030A
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parts
sodium
cell culture
culture medium
wide spectrum
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王文峰
曹先艳
马莹
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Dong Xun Bio Tech Ltd Granary
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Dong Xun Bio Tech Ltd Granary
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Abstract

The invention provides a kind of wide spectrum cell culture medium, including glucose, sodium acid carbonate, Sodium Pyruvate, sericin, potassium chloride, anhydrous magnesium sulfate, sodium chloride, AMSP, folic acid, inositol, niacinamide, anhydrous calcium chloride, ferric nitrate, succinic acid, sodium succinate, D calcium pantothenates, choline tartrate, riboflavin, thiamine hydrochloride, pyridoxine hydrochloride, N N-isopropylacrylamides, leukotrienes, monoethanolamine, aromatic esters, phenol red sodium and deionized water.Wide spectrum cell culture medium of the present invention, its component is rationally, nutritious, and more nutrients can be provided for cell culture（Including growth factor etc.）, high cell growth speed, cell state is good, sharpness of border, and Microscopic observation is bright, split coil method is more, form and decentralization are good, can be conducive to biological with healthy correlative study.

Description

A kind of wide spectrum cell culture medium
Technical field
The invention belongs to biological technology application, in particular it relates to a kind of wide spectrum cell culture medium.
Background technology
Cell culture is a biological important technology with healthy Related Research Domain.With the rapid hair of life science Exhibition, current cell culture turns into the important base of the disciplinary studies such as cell biology, molecular biology, science of heredity and immunology Plinth.In order to further investigated cell vegetative activity rule, have the pathology of related disorders and the pharmacology of medicine（Toxicity）Mechanism, exploitation tool There are different targetedly cell culture processes significant.
Cell culture medium is the nutriment for cultivating cell, is wide variety of crucial skill in life and regenerative medicine field Art.Especially as the rise of cell therapy personalization technology, the cell of amplification in vitro special efficacy into preclinical study and The emphasis development project of clinical treatment.Cell with amplification in vitro special efficacy includes thin by multi-functional candidate stem cell, T Born of the same parents, NK cells, DC cells and CIK cell etc., will again feed back in lactation organism, favorably after this kind of cell expansion ex vivo In the hematopoietic device function of improving body：Such as lifted red blood cell, the quantity of immunocyte and function safeguard, anti-tumor capacity with And immune defense ability.
The wide spectrum cell culture medium of the present invention is nutritious, goes for the training of the various kinds of cell in people source and inhuman source Support.
The content of the invention
Goal of the invention：The invention provides a kind of wide spectrum cell culture medium, for a variety of people sources and non-human archeocyte Culture.
Technical scheme：The invention provides a kind of wide spectrum cell culture medium, by weight part, including following components：Portugal 80-90 parts of grape sugar, 10-20 parts of sodium acid carbonate, 8-18 parts of Sodium Pyruvate, 3-12 parts of sericin, 30-50 parts of potassium chloride, nothing 5-12 parts of water magnesium sulfate, 60-80 parts of sodium chloride, 8-16 parts of AMSP, 0.2-0.6 parts of folic acid, inositol 0.5- 1.5 parts, 0.2-0.8 parts of niacinamide, 20-40 parts of anhydrous calcium chloride, 0.1-0.5 parts of ferric nitrate, 5-10 parts of succinic acid, succinic acid 9-18 parts of sodium, 0.2-0.8 parts of D-VB5 calcium, 0.5-1 parts of choline tartrate, 0.1-0.3 parts of riboflavin, thiamine hydrochloride 0.1-0.5 Part, 0.1-0.6 parts of pyridoxine hydrochloride, 0.1-0.5 parts of NIPA, 0.3-3 parts of leukotrienes, monoethanolamine 0.5-0.8 Part, 0.2-0.9 parts of aromatic esters, phenol red sodium 0.8-1.5 parts and 100 parts of deionized water.
Further, above-mentioned wide spectrum cell culture medium, 82-88 parts of glucose, 12-25 parts of sodium acid carbonate, Sodium Pyruvate 10-16 parts, 5-9 parts of sericin, 35-45 parts of potassium chloride, 6-10 parts of anhydrous magnesium sulfate, 70-78 parts of sodium chloride, anhydrous phosphoric acid 10-14 parts of sodium dihydrogen, 0.3-0.5 parts of folic acid, 0.8-1.2 parts of inositol, 0.3-0.6 parts of niacinamide, 25-35 parts of anhydrous calcium chloride, 0.2-0.4 parts of ferric nitrate, 6-8 parts of succinic acid, 12-16 parts of sodium succinate, 0.3-0.6 parts of D-VB5 calcium, choline tartrate 0.6- 0.8 part, 0.1-0.3 parts of riboflavin, 0.2-0.4 parts of thiamine hydrochloride, 0.3-0.5 parts of pyridoxine hydrochloride, NIPA 0.2-0.4 parts, 1-2 parts of leukotrienes, 0.6-0.8 parts of monoethanolamine, 0.3-0.8 parts of aromatic esters, phenol red sodium 1-1.3 parts and deionization 100 parts of water.
It is used as a kind of preferred embodiment of the present invention, above-mentioned wide spectrum cell culture medium, 86 parts of glucose, sodium acid carbonate 18 Part, 12 parts of Sodium Pyruvate, 6 parts of sericin, 40 parts of potassium chloride, 8 parts of anhydrous magnesium sulfate, 75 parts of sodium chloride, anhydrous phosphoric acid dihydro 12 parts of sodium, 0.4 part of folic acid, 1 part of inositol, 0.5 part of niacinamide, 30 parts of anhydrous calcium chloride, 0.3 part of ferric nitrate, 7 parts of succinic acid, fourth 14 parts of diacid sodium, 0.5 part of D-VB5 calcium, 0.7 part of choline tartrate, 0.2 part of riboflavin, 0.3 part of thiamine hydrochloride, hydrochloric acid pyrrole are trembled Pungent 0.4 part, 0.3 part of NIPA, 1.6 parts of leukotrienes, 0.7 part of monoethanolamine, 0.5 part of aromatic esters, phenol red sodium 1.2 parts and 100 parts of deionized water.
Further, above-mentioned wide spectrum cell culture medium, in addition to 3-8 parts of amino acid, the amino acid is with components by weight percent Meter, it is composed of the following components：5-10 parts of L- R-genes, 3-8 parts of L- hydrochloric acid cystines, 3-6 parts of Serine, glycine 1-5 parts, 4-6 parts of L- histidine monohydrochlorides, 8-20 parts of ILE, 7-18 parts of L-Leu, 10-20 parts of LYS, 1-6 parts of METHIONINE, 2-8 parts of L-phenylalanine, 5-15 parts of L-threonine, 1-3 parts of L-Trp, 5-9 parts of TYR and 8-12 parts of Valine.
Further, above-mentioned wide spectrum cell culture medium, in addition to 2-5 parts of confactors, the confactor is with weight Component meter, it is composed of the following components：2-10 parts of recombination human basic fibroblast growth factor, 3-8 parts of interleukin-22, interleukin 1-4 parts of 4 3-6 parts, 1-5 parts of interleukin-11 4,3-6 parts of IFN-γ and neuroleukin.
Further, above-mentioned wide spectrum cell culture medium, includes volume ratio 10-30% hyclone.Can be further For the cells with nutrient material of culture.
Further, above-mentioned wide spectrum cell culture medium, by weight part, in addition to 0.006 part of penicillin and 0.01 The streptomysin of part.Culture cell can be effectively prevented to be contaminated.
Further, above-mentioned wide spectrum cell culture medium, the penicillin is selected from 10000U/ml, and the streptomysin is selected from 10000μg/ml.Penicillin and streptomysin activity are good, and effect is good.
Beneficial effect：Wide spectrum cell culture medium of the present invention, its component is rationally, nutritious, can be cell training Support and more nutrients are provided（Including growth factor etc.）, high cell growth speed, cell state is good, sharpness of border, Jing Xiaguan Examine more bright, split coil method, form and decentralization good, biological and healthy correlative study can be conducive to.
Embodiment
Below will be by several specific embodiments, the present invention is furture elucidated, these embodiments simply to illustrate that problem, It is not a kind of limitation.
Embodiment 1
A kind of wide spectrum cell culture medium, by weight part, including following components：80 parts of glucose, 10 parts of sodium acid carbonate, third 8 parts of ketone acid sodium, 3 parts of sericin, 30 parts of potassium chloride, 5 parts of anhydrous magnesium sulfate, 60 parts of sodium chloride, AMSP 8 Part, 0.2 part of folic acid, 0.5 part of inositol, 0.2 part of niacinamide, 20 parts of anhydrous calcium chloride, 0.1 part of ferric nitrate, 5 parts of succinic acid, fourth 9 parts of diacid sodium, 0.2 part of D-VB5 calcium, 0.5 part of choline tartrate, 0.1 part of riboflavin, 0.1 part of thiamine hydrochloride, pyridoxine hydrochloride 0.1 part, 0.1 part of NIPA, 0.3 part of leukotrienes, 0.5 part of monoethanolamine, 0.2 part of aromatic esters, 0.8 part of phenol red sodium With 100 parts of deionized water.
In addition, above-mentioned wide spectrum cell culture medium also includes 3 parts of amino acid, the amino acid by weight part, by following Component is constituted：5 parts of L- R-genes, 3 parts of L- hydrochloric acid cystine, 3 parts of Serine, 1 part of glycine, L- histidine monohydrochlorides 4 parts, 8 parts of ILE, 7 parts of L-Leu, 10 parts of LYS, 1 part of METHIONINE, 2 parts of L-phenylalanine, L- 8 parts of 5 parts of threonine, 1 part of L-Trp, 5 parts of TYR and Valine.
Again, above-mentioned wide spectrum cell culture medium also includes 2 parts of confactors, the confactor by weight part, by Following components is constituted：2 parts of recombination human basic fibroblast growth factor, 3 parts of interleukin-22,3 parts of IL-4, interleukin-11 4 1 part of 1 part, 3 parts of IFN-γ and neuroleukin.
In addition, above-mentioned wide spectrum cell culture medium, includes the hyclone of volume ratio 10%.Also, with components by weight percent Meter, in addition to 0.006 part of penicillin and 0.01 part of streptomysin.Wherein, the penicillin is selected from 10000U/ml, the strepto- Element is selected from 10000 μ g/ml.
Embodiment 2
A kind of wide spectrum cell culture medium, by weight part, including following components：90 parts of glucose, 20 parts of sodium acid carbonate, acetone Sour 18 parts of sodium, 12 parts of sericin, 50 parts of potassium chloride, 12 parts of anhydrous magnesium sulfate, 80 parts of sodium chloride, AMSP 16 Part, 0.6 part of folic acid, 1.5 parts of inositol, 0.8 part of niacinamide, 40 parts of anhydrous calcium chloride, 0.5 part of ferric nitrate, 10 parts of succinic acid, fourth 18 parts of diacid sodium, 0.8 part of D-VB5 calcium, 1 part of choline tartrate, 0.3 part of riboflavin, 0.5 part of thiamine hydrochloride, pyridoxine hydrochloride 0.6 part, 0.5 part of NIPA, 3 parts of leukotrienes, 0.8 part of monoethanolamine, 0.9 part of aromatic esters, 1.5 parts of phenol red sodium and 100 parts of deionized water.
In addition, above-mentioned wide spectrum cell culture medium also includes 8 parts of amino acid, the amino acid by weight part, by following Component is constituted：10 parts of L- R-genes, 8 parts of L- hydrochloric acid cystine, 6 parts of Serine, 5 parts of glycine, L- hydrochloric acid group ammonia 6 parts of acid, 20 parts of ILE, 18 parts of L-Leu, 20 parts of LYS, 6 parts of METHIONINE, L-phenylalanine 8 Part, 15 parts of L-threonine, 3 parts of L-Trp, 9 parts of TYR and 12 parts of Valine.
Again, above-mentioned wide spectrum cell culture medium also includes 5 parts of confactors, the confactor by weight part, by Following components is constituted：10 parts of recombination human basic fibroblast growth factor, 8 parts of interleukin-22,6 parts of IL-4, interleukin 4 parts of 14 5 parts, 6 parts of IFN-γ and neuroleukin.
In addition, above-mentioned wide spectrum cell culture medium, includes the hyclone of volume ratio 30%.Also, with components by weight percent Meter, in addition to 0.006 part of penicillin and 0.01 part of streptomysin.Wherein, the penicillin is selected from 10000U/ml, the strepto- Element is selected from 10000 μ g/ml.
Embodiment 3
A kind of wide spectrum cell culture medium, by weight part, including following components：82 parts of glucose, 12 parts of sodium acid carbonate, third 10 parts of ketone acid sodium, 5 parts of sericin, 35 parts of potassium chloride, 6 parts of anhydrous magnesium sulfate, 70 parts of sodium chloride, AMSP 10 Part, 0.3 part of folic acid, 0.8 part of inositol, 0.3 part of niacinamide, 25 parts of anhydrous calcium chloride, 0.2 part of ferric nitrate, 6 parts of succinic acid, fourth two Sour 12 parts of sodium, 0.3 part of D-VB5 calcium, 0.6 part of choline tartrate, 0.1 part of riboflavin, 0.2 part of thiamine hydrochloride, pyridoxine hydrochloride 0.3 part, 0.2 part of NIPA, 1 part of leukotrienes, 0.6 part of monoethanolamine, 0.3 part of aromatic esters, 1 part of phenol red sodium and go 100 parts of ionized water.
In addition, above-mentioned wide spectrum cell culture medium also includes 3 parts of amino acid, the amino acid by weight part, by following Component is constituted：5 parts of L- R-genes, 3 parts of L- hydrochloric acid cystine, 3 parts of Serine, 1 part of glycine, L- histidine monohydrochlorides 4 parts, 8 parts of ILE, 7 parts of L-Leu, 10 parts of LYS, 1 part of METHIONINE, 2 parts of L-phenylalanine, L- 8 parts of 5 parts of threonine, 1 part of L-Trp, 5 parts of TYR and Valine.
Again, above-mentioned wide spectrum cell culture medium also includes 2 parts of confactors, the confactor by weight part, by Following components is constituted：2 parts of recombination human basic fibroblast growth factor, 3 parts of interleukin-22,3 parts of IL-4, interleukin-11 4 1 part of 1 part, 3 parts of IFN-γ and neuroleukin.
In addition, above-mentioned wide spectrum cell culture medium, includes the hyclone of volume ratio 10%.Also, with components by weight percent Meter, in addition to 0.006 part of penicillin and 0.01 part of streptomysin.Wherein, the penicillin is selected from 10000U/ml, the strepto- Element is selected from 10000 μ g/ml.
Embodiment 4
A kind of wide spectrum cell culture medium, by weight part, including following components：88 parts of glucose, 25 parts of sodium acid carbonate, third 16 parts of ketone acid sodium, 9 parts of sericin, 45 parts of potassium chloride, 10 parts of anhydrous magnesium sulfate, 78 parts of sodium chloride, AMSP 14 Part, 0.5 part of folic acid, 1.2 parts of inositol, 0.6 part of niacinamide, 35 parts of anhydrous calcium chloride, 0.4 part of ferric nitrate, 8 parts of succinic acid, fourth two Sour 16 parts of sodium, 0.6 part of D-VB5 calcium, 0.8 part of choline tartrate, 0.3 part of riboflavin, 0.4 part of thiamine hydrochloride, pyridoxine hydrochloride 0.5 part, 0.4 part of NIPA, 2 parts of leukotrienes, 0.8 part of monoethanolamine, 0.8 part of aromatic esters, 1.3 parts of phenol red sodium and 100 parts of deionized water.
In addition, above-mentioned wide spectrum cell culture medium also includes 8 parts of amino acid, the amino acid by weight part, by following Component is constituted：10 parts of L- R-genes, 8 parts of L- hydrochloric acid cystine, 6 parts of Serine, 5 parts of glycine, L- hydrochloric acid group ammonia 6 parts of acid, 20 parts of ILE, 18 parts of L-Leu, 20 parts of LYS, 6 parts of METHIONINE, L-phenylalanine 8 Part, 15 parts of L-threonine, 3 parts of L-Trp, 9 parts of TYR and 12 parts of Valine.
Again, above-mentioned wide spectrum cell culture medium also includes 5 parts of confactors, the confactor by weight part, by Following components is constituted：8 parts of recombination human basic fibroblast growth factor, 8 parts of interleukin-22,4 parts of IL-4, interleukin-11 4 4 parts of 5 parts, 6 parts of IFN-γ and neuroleukin.
In addition, above-mentioned wide spectrum cell culture medium, includes the hyclone of volume ratio 30%.Also, with components by weight percent Meter, in addition to 0.006 part of penicillin and 0.01 part of streptomysin.Wherein, the penicillin is selected from 10000U/ml, the strepto- Element is selected from 10000 μ g/ml.
Embodiment 5
A kind of wide spectrum cell culture medium, by weight part, including following components：86 parts of glucose, 18 parts of sodium acid carbonate, third 12 parts of ketone acid sodium, 6 parts of sericin, 40 parts of potassium chloride, 8 parts of anhydrous magnesium sulfate, 75 parts of sodium chloride, AMSP 12 Part, 0.4 part of folic acid, 1 part of inositol, 0.5 part of niacinamide, 30 parts of anhydrous calcium chloride, 0.3 part of ferric nitrate, 7 parts of succinic acid, succinic acid 14 parts of sodium, 0.5 part of D-VB5 calcium, 0.7 part of choline tartrate, 0.2 part of riboflavin, 0.3 part of thiamine hydrochloride, pyridoxine hydrochloride 0.4 part, 0.3 part of NIPA, 1.6 parts of leukotrienes, 0.7 part of monoethanolamine, 0.5 part of aromatic esters, 1.2 parts of phenol red sodium With 100 parts of deionized water.
In addition, above-mentioned wide spectrum cell culture medium also includes 6 parts of amino acid, the amino acid by weight part, by following Component is constituted：8 parts of L- R-genes, 6 parts of L- hydrochloric acid cystine, 5 parts of Serine, 3 parts of glycine, L- histidine monohydrochlorides 5 parts, 12 parts of ILE, 10 parts of L-Leu, 15 parts of LYS, 3 parts of METHIONINE, 4 parts of L-phenylalanine, 9 parts of 9 parts of L-threonine, 2 parts of L-Trp, 6 parts of TYR and Valine.
Again, above-mentioned wide spectrum cell culture medium also includes 4 parts of confactors, the confactor by weight part, by Following components is constituted：6 parts of recombination human basic fibroblast growth factor, 5 parts of interleukin-22,4 parts of IL-4, interleukin-11 4 3 parts of 5 parts, 5 parts of IFN-γ and neuroleukin.
In addition, above-mentioned wide spectrum cell culture medium, includes the hyclone of volume ratio 20%.Also, with components by weight percent Meter, in addition to 0.006 part of penicillin and 0.01 part of streptomysin.Wherein, the penicillin is selected from 10000U/ml, the strepto- Element is selected from 10000 μ g/ml.
Described above is only several embodiments of invention, it is noted that for those skilled in the art For, on the premise of inventive principle is not departed from, some improvement can also be made, these improvement also should be regarded as the protection of the present invention Scope.

Claims (8)

1. a kind of wide spectrum cell culture medium, it is characterised in that：By weight part, including following components：80-90 parts of glucose, 10-20 parts of sodium acid carbonate, 8-18 parts of Sodium Pyruvate, 3-12 parts of sericin, 30-50 parts of potassium chloride, anhydrous magnesium sulfate 5-12 Part, 60-80 parts of sodium chloride, 8-16 parts of AMSP, 0.2-0.6 parts of folic acid, 0.5-1.5 parts of inositol, niacinamide 0.2-0.8 parts, 20-40 parts of anhydrous calcium chloride, 0.1-0.5 parts of ferric nitrate, 5-10 parts of succinic acid, 9-18 parts of sodium succinate, D- it is general Sour calcium 0.2-0.8 parts, 0.5-1 parts of choline tartrate, 0.1-0.3 parts of riboflavin, 0.1-0.5 parts of thiamine hydrochloride, pyridoxine hydrochloride 0.1-0.6 parts, 0.1-0.5 parts of NIPA, 0.3-3 parts of leukotrienes, 0.5-0.8 parts of monoethanolamine, aromatic esters 0.2-0.9 parts, phenol red sodium 0.8-1.5 parts and 100 parts of deionized water.
2. wide spectrum cell culture medium according to claim 1, it is characterised in that：By weight part, including following components： 82-88 parts of glucose, 12-25 parts of sodium acid carbonate, 10-16 parts of Sodium Pyruvate, 5-9 parts of sericin, 35-45 parts of potassium chloride, nothing 6-10 parts of water magnesium sulfate, 70-78 parts of sodium chloride, 10-14 parts of AMSP, 0.3-0.5 parts of folic acid, inositol 0.8- 1.2 parts, 0.3-0.6 parts of niacinamide, 25-35 parts of anhydrous calcium chloride, 0.2-0.4 parts of ferric nitrate, 6-8 parts of succinic acid, sodium succinate 12-16 parts, 0.3-0.6 parts of D-VB5 calcium, 0.6-0.8 parts of choline tartrate, 0.1-0.3 parts of riboflavin, thiamine hydrochloride 0.2- 0.4 part, 0.3-0.5 parts of pyridoxine hydrochloride, 0.2-0.4 parts of NIPA, 1-2 parts of leukotrienes, monoethanolamine 0.6- 0.8 part, 0.3-0.8 parts of aromatic esters, phenol red sodium 1-1.3 parts and 100 parts of deionized water.
3. wide spectrum cell culture medium according to claim 1, it is characterised in that：By weight part, including following components： 86 parts of glucose, 18 parts of sodium acid carbonate, 12 parts of Sodium Pyruvate, 6 parts of sericin, 40 parts of potassium chloride, 8 parts of anhydrous magnesium sulfate, chlorine Change 75 parts of sodium, 12 parts of AMSP, 0.4 part of folic acid, 1 part of inositol, 0.5 part of niacinamide, 30 parts of anhydrous calcium chloride, nitre Sour 0.3 part of iron, 7 parts of succinic acid, 14 parts of sodium succinate, 0.5 part of D-VB5 calcium, 0.7 part of choline tartrate, 0.2 part of riboflavin, 0.3 part of thiamine hydrochloride, 0.4 part of pyridoxine hydrochloride, 0.3 part of NIPA, 1.6 parts of leukotrienes, 0.7 part of monoethanolamine, 100 parts of 0.5 part of aromatic esters, 1.2 parts of phenol red sodium and deionized water.
4. the wide spectrum cell culture medium according to claim any one of 1-3, it is characterised in that：Also include 3-8 parts of amino acid, The amino acid is by weight part, composed of the following components：5-10 parts of L- R-genes, 3-8 parts of L- hydrochloric acid cystines, 3-6 parts of Serine, 1-5 parts of glycine, 4-6 parts of L- histidine monohydrochlorides, 8-20 parts of ILE, 7-18 parts of L-Leu, 10-20 parts of LYS, 1-6 parts of METHIONINE, 2-8 parts of L-phenylalanine, 5-15 parts of L-threonine, L-Trp 1- 8-12 parts of 3 parts, 5-9 parts of TYR and Valine.
5. wide spectrum cell culture medium according to claim 4, it is characterised in that：Also include 2-5 parts of confactors, it is described auxiliary Help the factor by weight part, it is composed of the following components：2-10 parts of recombination human basic fibroblast growth factor, interleukin-22 1-4 parts of 3-8 parts, 3-6 parts of IL-4,1-5 parts of interleukin-11 4,3-6 parts of IFN-γ and neuroleukin.
6. wide spectrum cell culture medium according to claim 5, it is characterised in that：Also include volume ratio 10-30% tire ox blood Clearly.
7. wide spectrum cell culture medium according to claim 6, it is characterised in that：By weight part, in addition to 0.006 part Penicillin and 0.01 part of streptomysin.
8. wide spectrum cell culture medium according to claim 7, it is characterised in that：The penicillin is selected from 10000U/ml, institute State streptomysin and be selected from 10000 μ g/ml.
CN201710525237.5A 2017-06-30 2017-06-30 A kind of wide spectrum cell culture medium Pending CN107217030A (en)

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