CN107164327A - A kind of method that breast cancer cells of MDA MB 231 expand culture - Google Patents

A kind of method that breast cancer cells of MDA MB 231 expand culture Download PDF

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CN107164327A
CN107164327A CN201710524260.2A CN201710524260A CN107164327A CN 107164327 A CN107164327 A CN 107164327A CN 201710524260 A CN201710524260 A CN 201710524260A CN 107164327 A CN107164327 A CN 107164327A
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mda
breast cancer
culture
cancer cells
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曹先艳
王文峰
施亮
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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 method that breast cancer cells of MDA MB 231 expand culture, the breast cancer cell special culture medias of MDA MB 231 are exhausted with liquid-transfering gun, add 3 5ml PBS washings cell 23 times, 1 2ml 0.38% trypsase is added into Tissue Culture Dish, 3 5min are digested under the conditions of 37 DEG C, the 2 3ml fresh breast cancer cell special culture medias of MDA MB 231 are added, Tissue Culture Dish bottom is blown and beaten 40 50 times using liquid-transfering gun；Liquid is moved in cell centrifuge tube, 3 10min are centrifuged under the conditions of 1000 1500g/min, supernatant is removed with liquid-transfering gun, the fresh breast cancer cell special culture medias of MDA MB 231 of 2 3ml are added, piping and druming cell 40 50 times above and below liquid-transfering gun are used；Averagely it is added in 38 Tissue Culture Dish, is enlarged culture.

Description

A kind of method that MDA-MB-231 breast cancer cells expand culture
Technical field
The invention belongs to biological technology application, in particular it relates to which a kind of MDA-MB-231 breast cancer cells expand training Foster method.
Background technology
Breast cancer is the most common cancer of women, mainly includes duct carcinoma and lobular carcinoma.U.S. CA in 2016（A Cancer Journal for Clinicians）The recent statistics data display of announcement, the U.S. is expected will there is 420840 women in 2016 Suffer from breast cancer, account for the 38% of women de novo malignancy, occupy the female malignant incidence of disease first, and death toll will reach 52930.More seriously global breast cancer incidence increases year by year.
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.Wherein, MDA-MB-231 breast cancer cells are high as humanized Metastatic breast cancer cell, is usually used in the in vivo and in vitro of breast cancer.
The content of the invention
Goal of the invention：The invention provides a kind of method that MDA-MB-231 breast cancer cells expand culture, for a variety of People source and the culture of non-human archeocyte.
Technical scheme：Expand the method cultivated the invention provides a kind of MDA-MB-231 breast cancer cells, including it is following Step：MDA-MB-231 breast cancer cells special culture media is exhausted with liquid-transfering gun, 3-5ml PBS washings cell is added 2-3 times, 1-2ml 0.38% trypsase is added into Tissue Culture Dish, 3-5min is digested under the conditions of 37 DEG C, the new of 2-3ml is added Fresh MDA-MB-231 breast cancer cell special culture medias, Tissue Culture Dish bottom is blown and beaten 40-50 times using liquid-transfering gun；Liquid is moved Into cell centrifuge tube, 3-10min is centrifuged under the conditions of 1000-1500g/min, supernatant is removed with liquid-transfering gun, 2-3ml is added Fresh MDA-MB-231 breast cancer cell special culture medias, use piping and druming cell 40-50 times above and below liquid-transfering gun；Averagely it is added to In 3-8 Tissue Culture Dish, culture is enlarged.MDA-MB-231 breast cancer cells of the present invention expand the side of culture Method, method is reasonable, it is easy to accomplish, high cell growth speed, cell state is good, sharpness of border, Microscopic observation is bright, split coil method more Many, form and decentralization are good, can conveniently be applied to the in vivo and in vitro of breast cancer.
Further, above-mentioned MDA-MB-231 breast cancer cells expand also wraps in the method for culture, the trypsase Containing 0.01% EDTA.It can be engaged with trypsase, attached cell is departed from culture dish rapidly, reduce trypsase to thin The damage of born of the same parents.
Further, above-mentioned MDA-MB-231 breast cancer cells expand the method for culture, the MDA-MB-231 mammary gland The hyclone of cancer cell special basal medium and 10-30% including 70-90%.Hyclone can be further culture Cells with nutrient material, while the digestion of trypsase can also be terminated.
Further, above-mentioned MDA-MB-231 breast cancer cells expand the method for culture, and the basal medium is with weight Measure component meter, including following components：80-90 parts of glucose, 10-20 parts of sodium acid carbonate, 8-18 parts of Sodium Pyruvate, aliphatic acid are white 4-16 parts of albumen, 30-50 parts of potassium chloride, 5-12 parts of anhydrous magnesium sulfate, 60-80 parts of sodium chloride, AMSP 8-16 Part, 0.2-0.6 parts of folic acid, 0.5-1.5 parts of inositol, 0.2-0.8 parts of niacinamide, 20-40 parts of anhydrous calcium chloride, ferric nitrate 0.1- 0.5 part, 5-10 parts of succinic acid, 9-18 parts of sodium succinate, 0.2-0.8 parts of D-VB5 calcium, N- isopropyl acrylamides 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 part, 0.2-1 parts of L- propionamide-L- paddy ammonia dipeptides, 1-3 parts of acetic acid esters, 0.5-0.8 parts of biotin, 0.2-0.8 parts of lipoic acid, Phenol red sodium 0.8-1.5 parts and 100 parts of deionized water.
Further, above-mentioned MDA-MB-231 breast cancer cells expand the method for culture, and the basal medium is also wrapped 3-8 parts of amino acid are included, the amino acid is by weight part, composed of the following components：5-10 parts of L- R-genes, L- salt Sour cystine 3-8 parts, 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, L-threonine 5- 8-12 parts of 15 parts, 1-3 parts of L-Trp, 5-9 parts of TYR and Valine.
Further, above-mentioned MDA-MB-231 breast cancer cells expand the method for culture, and the basal medium is also wrapped 2-5 parts of confactors are included, the confactor is by weight part, composed of the following components：3-8 parts of insulin-like growth factor, 2-5 parts of interleukin 6,1-4 parts of IL-10,3-6 parts of interleukin 12,2-10 parts, GM-CSF1-5 parts of TNF-β.Basis culture Base component is rationally, nutritious, and more nutrients can be provided for cell culture.
Further, above-mentioned MDA-MB-231 breast cancer cells expand the method for culture, and the basal medium is with weight Measure component meter, in addition to 0.006 part of penicillin and 0.01 part of streptomysin.The streptomysin is selected from 10000 μ g/ml.Can have Effect prevents culture cell to be contaminated.
Further, above-mentioned MDA-MB-231 breast cancer cells expand the method for culture, and the penicillin is selected from 10000U/ml, penicillin and streptomysin activity are good, and effect is good.
Further, above-mentioned MDA-MB-231 breast cancer cells expand the method for culture, and the Tissue Culture Dish is 10cm Tissue Culture Dish.
Beneficial effect：The method that MDA-MB-231 breast cancer cells of the present invention expand culture, method is reasonable, it is easy to Realize, high cell growth speed, cell state is good, wherein, MDA-MB-231 breast cancer cells special culture media can be cell Culture provides more nutrients（Including growth factor etc.）, high cell growth speed, cell state is good, sharpness of border, under mirror Observe more bright, split coil method, form and decentralization more preferably, 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 method that MDA-MB-231 breast cancer cells expand culture, comprises the following steps：With liquid-transfering gun by MDA-MB-231 Breast cancer cell special culture media is exhausted, and adds 3ml PBS washings cell 2 times, and 1ml is added into 10cm Tissue Culture Dish 0.38% trypsase, digests 5min under the conditions of 37 DEG C, and the fresh MDA-MB-231 breast cancer cells for adding 2ml are special Culture medium, Tissue Culture Dish bottom is blown and beaten 40 times using liquid-transfering gun；Liquid is moved in cell centrifuge tube, in 1000g/min conditions Lower centrifugation 10min, supernatant is removed with liquid-transfering gun, is added the fresh MDA-MB-231 breast cancer cell special culture medias of 2ml, is made With piping and druming cell 40 times above and below liquid-transfering gun；Averagely it is added in 3 Tissue Culture Dish, is enlarged culture.
Wherein, the MDA-MB-231 breast cancer cells special culture media includes 70% basal medium and 30% tire Cow's serum.The basal medium by weight part, including following components：80 parts of glucose, 10 parts of sodium acid carbonate, acetone Sour 8 parts of sodium, fatty 4 parts of syntonin, 30 parts of potassium chloride, 5 parts of anhydrous magnesium sulfate, 60 parts of sodium chloride, AMSP 8 parts, 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, 0.2 part of N- isopropyl acrylamides, 0.5 part of choline tartrate, riboflavin 0.1 Part, 0.1 part of thiamine hydrochloride, 0.1 part of pyridoxine hydrochloride, 0.2 part of L- propionamide-L- paddy ammonia dipeptides, 1 part of acetic acid esters, biotin 100 parts of 0.5 part, 0.2 part of lipoic acid, 0.8 part of phenol red sodium and deionized water.
In addition, the basal medium also includes 3 parts of amino acid, the amino acid by weight part, by following components group Into：5 parts of L- R-genes, 3 parts of L- hydrochloric acid cystine, 3 parts of Serine, 1 part of glycine, 4 parts of L- histidine monohydrochlorides, 8 parts of ILE, 7 parts of L-Leu, 10 parts of LYS, 1 part of METHIONINE, 2 parts of L-phenylalanine, L- Soviet Unions ammonia 8 parts of 5 parts of acid, 1 part of L-Trp, 5 parts of TYR and Valine.
Again, the basal medium also includes 2 parts of confactors, the confactor by weight part, by with the following group It is grouped into：3 parts of insulin-like growth factor, 2 parts of interleukin 6,1 part of IL-10,3 parts of interleukin 12,2 parts of TNF-β, GM- CSF1 parts.
Further, the basal medium by weight part, in addition to 0.006 part of penicillin and 0.01 part of strepto- Element.Also, the penicillin is selected from 10000U/ml, the streptomysin is selected from 10000 μ g/ml.In addition, in the trypsase Also include 0.01% EDTA.
Embodiment 2
A kind of method that MDA-MB-231 breast cancer cells expand culture, comprises the following steps：With liquid-transfering gun by MDA-MB-231 Breast cancer cell special culture media is exhausted, and adds 5ml PBS washings cell 3 times, and 2ml is added into 10cm Tissue Culture Dish 0.38% trypsase, digests 3min under the conditions of 37 DEG C, and the fresh MDA-MB-231 breast cancer cells for adding 3ml are special Culture medium, Tissue Culture Dish bottom is blown and beaten 50 times using liquid-transfering gun；Liquid is moved in cell centrifuge tube, in 1500g/min conditions Lower centrifugation 3min, supernatant is removed with liquid-transfering gun, is added the fresh MDA-MB-231 breast cancer cell special culture medias of 3ml, is made With piping and druming cell 50 times above and below liquid-transfering gun；Averagely it is added in 8 Tissue Culture Dish, is enlarged culture.
Wherein, the MDA-MB-231 breast cancer cells special culture media includes 90% basal medium and 10% tire Cow's serum.The basal medium by weight part, including following components：90 parts of glucose, 20 parts of sodium acid carbonate, acetone Sour 18 parts of sodium, fatty 16 parts of syntonin, 50 parts of potassium chloride, 12 parts of anhydrous magnesium sulfate, 80 parts of sodium chloride, anhydrous phosphoric acid dihydro 16 parts of sodium, 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, succinic acid 10 Part, 18 parts of sodium succinate, 0.8 part of D-VB5 calcium, 0.8 part of N- isopropyl acrylamides, 1 part of choline tartrate, riboflavin 0.3 part, 0.5 part of thiamine hydrochloride, 0.6 part of pyridoxine hydrochloride, 1 part of L- propionamide-L- paddy ammonia dipeptides, 3 parts of acetic acid esters, biotin 100 parts of 0.8 part, 0.8 part of lipoic acid, 1.5 parts of phenol red sodium and deionized water.
In addition, the basal medium also includes 8 parts of amino acid, the amino acid by weight part, by following components group Into：10 parts of L- R-genes, 8 parts of L- hydrochloric acid cystine, 6 parts of Serine, 5 parts of glycine, 6 parts of L- histidine monohydrochlorides, 20 parts of ILE, 18 parts of L-Leu, 20 parts of LYS, 6 parts of METHIONINE, 8 parts of L-phenylalanine, L- Soviet Unions 12 parts of 15 parts of propylhomoserin, 3 parts of L-Trp, 9 parts of TYR and Valine.
Again, the basal medium also includes 5 parts of confactors, the confactor by weight part, by with the following group It is grouped into：8 parts of insulin-like growth factor, 5 parts of interleukin 6,4 parts of IL-10,6 parts of interleukin 12,10 parts of TNF-β, GM- 5 parts of CSF.
Further, the basal medium by weight part, in addition to 0.006 part of penicillin and 0.01 part of strepto- Element.Also, the penicillin is selected from 10000U/ml, the streptomysin is selected from 10000 μ g/ml.In addition, in the trypsase Also include 0.01% EDTA.
Embodiment 3
A kind of method that MDA-MB-231 breast cancer cells expand culture, comprises the following steps：With liquid-transfering gun by MDA-MB-231 Breast cancer cell special culture media is exhausted, and adds 4ml PBS washings cell 2 times, and 2ml is added into 10cm Tissue Culture Dish 0.38% trypsase, digests 4min under the conditions of 37 DEG C, and the fresh MDA-MB-231 breast cancer cells for adding 2ml are special Culture medium, Tissue Culture Dish bottom is blown and beaten 48 times using liquid-transfering gun；Liquid is moved in cell centrifuge tube, in 1200g/min conditions Lower centrifugation 5min, supernatant is removed with liquid-transfering gun, is added the fresh MDA-MB-231 breast cancer cell special culture medias of 3ml, is made With piping and druming cell 45 times above and below liquid-transfering gun；Averagely it is added in 6 Tissue Culture Dish, is enlarged culture.
Wherein, the MDA-MB-231 breast cancer cells special culture media includes 80% basal medium and 20% tire Cow's serum.The basal medium by weight part, including following components：86 parts of glucose, 18 parts of sodium acid carbonate, acetone Sour 12 parts of sodium, fatty 9 parts of syntonin, 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, 0.5 part of N- isopropyl acrylamides, 0.7 part of choline tartrate, 0.2 part of riboflavin, 0.3 part of thiamine hydrochloride, 0.4 part of pyridoxine hydrochloride, 0.6 part of L- propionamide-L- paddy ammonia dipeptides, 1.8 parts of acetic acid esters, biotin 0.7 Part, 0.6 part of lipoic acid, 1.2 parts of phenol red sodium and 100 parts of deionized water.
In addition, the basal medium also includes 5 parts of amino acid, the amino acid by weight part, by following components group Into：8 parts of L- R-genes, 6 parts of L- hydrochloric acid cystine, 5 parts of Serine, 3 parts of glycine, 5 parts of L- histidine monohydrochlorides, 12 parts of ILE, 10 parts of L-Leu, 15 parts of LYS, 3 parts of METHIONINE, 4 parts of L-phenylalanine, L- Soviet Unions 9 parts of 9 parts of propylhomoserin, 2 parts of L-Trp, 6 parts of TYR and Valine.
Again, the basal medium also includes 4 parts of confactors, the confactor by weight part, by with the following group It is grouped into：5 parts of insulin-like growth factor, 3 parts of interleukin 6,2 parts of IL-10,5 parts of interleukin 12,8 parts of TNF-β, GM- CSF2 parts.
Further, the basal medium by weight part, in addition to 0.006 part of penicillin and 0.01 part of strepto- Element.Also, the penicillin is selected from 10000U/ml, the streptomysin is selected from 10000 μ g/ml.In addition, in the trypsase Also include 0.01% EDTA.
Embodiment 4
A kind of method that MDA-MB-231 breast cancer cells expand culture, comprises the following steps：With liquid-transfering gun by MDA-MB-231 Breast cancer cell special culture media is exhausted, and adds 5ml PBS washings cell 2 times, and 2ml is added into 10cm Tissue Culture Dish 0.38% trypsase, digests 5min under the conditions of 37 DEG C, and the fresh MDA-MB-231 breast cancer cells for adding 2-3ml are special With culture medium, Tissue Culture Dish bottom is blown and beaten 50 times using liquid-transfering gun；Liquid is moved in cell centrifuge tube, in 1500g/min bars 5min is centrifuged under part, supernatant is removed with liquid-transfering gun, the fresh MDA-MB-231 breast cancer cell special culture medias of 2ml are added, Use piping and druming cell 50 times above and below liquid-transfering gun；Averagely it is added in 6 Tissue Culture Dish, is enlarged culture.
Wherein, the MDA-MB-231 breast cancer cells special culture media includes 70 basal medium and 30% tire Cow's serum.The basal medium by weight part, including following components：82 parts of glucose, 25 parts of sodium acid carbonate, acetone Sour 10 parts of sodium, fatty 8 parts of syntonin, 38 parts of potassium chloride, 8 parts of anhydrous magnesium sulfate, 70 parts of sodium chloride, AMSP 10 parts, 0.5 part of folic acid, 1.2 parts of inositol, 0.5 part of niacinamide, 30 parts of anhydrous calcium chloride, 0.4 part of ferric nitrate, 6 parts of succinic acid, fourth 12 parts of diacid sodium, 0.6 part of D-VB5 calcium, 0.5 part of N- isopropyl acrylamides, 0.7 part of choline tartrate, riboflavin 0.2 Part, 0.4 part of thiamine hydrochloride, 0.3 part of pyridoxine hydrochloride, 0.8 part of L- propionamide-L- paddy ammonia dipeptides, 1.8 parts of acetic acid esters, biotin 100 parts of 0.6 part, 0.7 part of lipoic acid, 1 part of phenol red sodium and deionized water.
In addition, the basal medium also includes 6 parts of amino acid, the amino acid by weight part, by following components group Into：5 parts of L- R-genes, 8 parts of L- hydrochloric acid cystine, 6 parts of Serine, 1 part of glycine, 4 parts of L- histidine monohydrochlorides, 12 parts of ILE, 12 parts of L-Leu, 20 parts of LYS, 1 part of METHIONINE, 2 parts of L-phenylalanine, L- Soviet Unions 10 parts of 5 parts of propylhomoserin, 2 parts of L-Trp, 6 parts of TYR and Valine.
Again, the basal medium also includes 2 parts of confactors, the confactor by weight part, by with the following group It is grouped into：3 parts of insulin-like growth factor, 5 parts of interleukin 6,4 parts of IL-10,3 parts of interleukin 12,5 parts of TNF-β, GM- CSF3 parts.
Further, the basal medium by weight part, in addition to 0.006 part of penicillin and 0.01 part of strepto- Element.Also, the penicillin is selected from 10000U/ml, the streptomysin is selected from 10000 μ g/ml.In addition, in the trypsase Also include 0.01% EDTA.
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 (9)

1. a kind of method that MDA-MB-231 breast cancer cells expand culture, it is characterised in that：Comprise the following steps：Use liquid-transfering gun MDA-MB-231 breast cancer cells special culture media is exhausted, 3-5ml PBS washings cell is added 2-3 times, to Tissue Culture Dish Middle addition 1-2ml 0.38% trypsase, digests 3-5min under the conditions of 37 DEG C, adds 2-3ml fresh MDA-MB- 231 breast cancer cell special culture medias, Tissue Culture Dish bottom is blown and beaten 40-50 times using liquid-transfering gun；Liquid is moved into cell centrifugation Guan Zhong, centrifuges 3-10min under the conditions of 1000-1500g/min, and supernatant is removed with liquid-transfering gun, adds the fresh MDA- of 2-3ml MB-231 breast cancer cell special culture medias, use piping and druming cell 40-50 times above and below liquid-transfering gun；Averagely it is added to 3-8 cell In culture dish, culture is enlarged.
2. the method that MDA-MB-231 breast cancer cells according to claim 1 expand culture, it is characterised in that：The pancreas Also included in protease 0.01% EDTA.
3. the method that MDA-MB-231 breast cancer cells according to claim 1 or 2 expand culture, it is characterised in that：Institute State the hyclone of MDA-MB-231 breast cancer cells special basal medium and 10-30% including 70-90%.
4. the method that MDA-MB-231 breast cancer cells according to claim 3 expand culture, it is characterised in that：The base Basal culture medium by weight part, including following components：80-90 parts of glucose, 10-20 parts of sodium acid carbonate, Sodium Pyruvate 8- 18 parts, fatty 4-16 parts of syntonin, 30-50 parts of potassium chloride, 5-12 parts of anhydrous magnesium sulfate, 60-80 parts of sodium chloride, anhydrous phosphorus 8-16 parts of acid dihydride sodium, 0.2-0.6 parts of folic acid, 0.5-1.5 parts of inositol, 0.2-0.8 parts of niacinamide, anhydrous calcium chloride 20-40 Part, 0.1-0.5 parts of ferric nitrate, 5-10 parts of succinic acid, 9-18 parts of sodium succinate, 0.2-0.8 parts of D-VB5 calcium, N- isopropyl first 0.2-0.8 parts of base acrylamide, 0.5-1 parts of choline tartrate, 0.1-0.3 parts of riboflavin, 0.1-0.5 parts of thiamine hydrochloride, hydrochloric acid 0.1-0.6 parts of Benadon, 0.2-1 parts of L- propionamide-L- paddy ammonia dipeptides, 1-3 parts of acetic acid esters, 0.5-0.8 parts of biotin, sulphur are pungent 100 parts of 0.8-1.5 parts of sour 0.2-0.8 parts, phenol red sodium and deionized water.
5. the method that MDA-MB-231 breast cancer cells according to claim 4 expand culture, it is characterised in that：The base Basal culture medium also includes 3-8 parts of amino acid, and the amino acid is by weight part, composed of the following components：L- R-genes 5-10 parts, 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, L- it is different bright 8-20 parts of propylhomoserin, 7-18 parts of L-Leu, 10-20 parts of LYS, 1-6 parts of METHIONINE, L-phenylalanine 2-8 Part, 5-15 parts of L-threonine, 1-3 parts of L-Trp, 5-9 parts of TYR and 8-12 parts of Valine.
6. the method that MDA-MB-231 breast cancer cells according to claim 5 expand culture, it is characterised in that：The base Basal culture medium also includes 2-5 parts of confactors, and the confactor is by weight part, composed of the following components：Insulin is given birth to Long factor 3-8 parts, 2-5 parts of interleukin 6,1-4 parts of IL-10,3-6 parts of interleukin 12,2-10 parts of TNF-β, GM-CSF1-5 Part.
7. the method that MDA-MB-231 breast cancer cells according to claim 6 expand culture, it is characterised in that：The base Basal culture medium by weight part, in addition to 0.006 part of penicillin and 0.01 part of streptomysin.
8. the method that MDA-MB-231 breast cancer cells according to claim 7 expand culture, it is characterised in that：The green grass or young crops Mycin is selected from 10000U/ml, and the streptomysin is selected from 10000 μ g/ml.
9. the method that MDA-MB-231 breast cancer cells according to claim 1 expand culture, it is characterised in that：It is described thin Born of the same parents' culture dish is 10cm Tissue Culture Dish.
CN201710524260.2A 2017-06-30 2017-06-30 A kind of method that breast cancer cells of MDA MB 231 expand culture Pending CN107164327A (en)

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