CN102634482B - Serum-free complete medium for mesenchymal stem cell - Google Patents
Serum-free complete medium for mesenchymal stem cell Download PDFInfo
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Abstract
The invention discloses a serum-free complete medium for mesenchymal stem cell, consisting of a basal medium and an additive component, wherein the basal medium is DMEM/F12 (dulbecco modified eagle medium), and the additive component consists of recombinant human insulin, human serum albumin, human transferrin, cholesterol, sodium selenite, ferric nitrate and glucan. The complete medium is free from serum, so that the defects that the serum-containing medium is unstable in serum batches, high in cell toxicity, abundant in heterologous protein and the like can be greatly avoided, and a good tool can be provided for the basic research and the clinic treatment of the mesenchymal stem cell; and the invention further achieves the effect which is approximate or equal to the serum-containing complete medium and the other serum-free complete mediums in the aspect of cell proliferation, so that the long-term subcultring of the cell can be supported.
Description
Technical field
The present invention relates to field of cell culture, be specifically related to a kind of serum-free perfect medium of mescenchymal stem cell.
Background technology
Mescenchymal stem cell is from mesoblastic adult multipotential stem cell, can separate from multiple tissues such as marrow, periosteum, synovial membrane, muscle, fat, has the potential that directed differentiation is the broad variety mature cell.In whole vital process, histoorgan still is environment change and be upset and all need mescenchymal stem cell constantly to replenish renewal in steady state, therefore mescenchymal stem cell becomes cell engineering and the desirable seed cell of organizational project, yet the comparatively small amt of mescenchymal stem cell in the tissue such as marrow, be difficult to satisfy the demand of clinical treatment and research aspect, therefore need increase in a large number to mescenchymal stem cell external.
At present, all added a certain proportion of animal serum in the conventional culture system of mescenchymal stem cell, commonly used is foetal calf serum.Serum is absolutely necessary in the cells in vitro culture system, and it provides somatomedin, in conjunction with albumen, and hormone, and provide protection is provided; Yet because serum composition complexity, and contain the factor or toxicant that some are unfavorable for the cell growth, these materials may influence result of study, and also may be because there be pathogenic agent in animal serum, and clinical application is constituted a threat to, in addition, serum batch between there are differences, cause cultivation results difference also to occur.Although existing a lot of serum free mediums also contain a spot of animal serum mostly at present.
Summary of the invention
One of purpose of the present invention is to solve the deficiencies in the prior art, and a kind of serum-free perfect medium of human mesenchymal stem cell vitro culture is provided, and overcomes instability between the serum batch that contains blood serum medium, defectives such as cytotoxicity and a large amount of heterologous proteins are arranged.
Another purpose of the present invention is external mescenchymal stem cell to be increased in a large number.
For achieving the above object, the present invention has taked following technical scheme:
A kind of serum-free perfect medium of mescenchymal stem cell, constituted by basic medium and interpolation composition, described basic medium is DMEM/F12, and described interpolation composition is made up of recombinant human insulin, human serum albumin, human transferrin, cholesterol, Sodium Selenite, iron nitrate and dextran.
Further technical scheme is recombinant human insulin 2～10mg/L in the described interpolation composition, human serum albumin 1～30g/L, human transferrin 1～10mg/L, cholesterol 10～50mg/L, Sodium Selenite 0.05～0.2mg/L, iron nitrate 1～10mg/L, dextran 10～100mg/L.
Further technical scheme is that described basic medium DMEM/F12 model comprises 12400-024, Gibco 884400.
Further technical scheme is, described serum-free perfect medium is used for the vitro culture mescenchymal stem cell, and described mescenchymal stem cell comprises mesenchymal stem cells MSCs, umbilical cord mesenchymal stem cells.
Compared with prior art, the present invention has following beneficial effect:
(1) substratum of the present invention does not contain serum, avoided greatly containing unstable between the serum batch of blood serum medium, defectives such as cytotoxicity and a large amount of heterologous proteins are arranged, for fundamental research and the clinical treatment of mescenchymal stem cell provides good instrument.
(2) the present invention is promoting to have reached the effect approaching or suitable with containing blood serum medium and other serum free mediums aspect the cell increment, but the cultivation of going down to posterity for a long time of sustenticular cell.
Description of drawings
Fig. 1 is the streaming DNA cycle detection synoptic diagram of embodiment 1.
Fig. 2 is the streaming DNA cycle detection synoptic diagram of embodiment 2.
Fig. 3 is the streaming DNA cycle detection synoptic diagram of embodiment 3.
Embodiment
Embodiment one
The preparation of 1L mesenchymal stem cell serum-free perfect medium: with product type be 12400-024 DMEM/F12 dry powder (packing: 10 bags of * 1L/ boxes) 1 bag, recombinant human insulin 7mg, human serum albumin 12g, human transferrin 1mg, cholesterol 25mg, Sodium Selenite 0.185mg, iron nitrate 4mg, dextran 50mg mixes, and adds injection water 800ml, stirring and dissolving, and regulate pH to 7.1 with the hydrochloric acid of 0.5mol/L, be settled to 1L, with the degerming of 0.22um membrane filtration, deposit for 4 ℃ behind the mixing.
BMSCs (mesenchymal stem cells MSCs) vitro culture: extract volunteer's marrow 10ml by hospital, equal-volume is laid on the lymphocyte separation medium of 1.073g/ml, and the centrifugal 30min of 2250rpm gets cloud mononuclearcell layer, washes 3 times with physiological saline; Again successively with 2000rpm, 1800rpm, the centrifugal supernatant liquor that goes of 1600rpm, each centrifugal five minutes; The cell that goes to obtain after the supernatant liquor is resuspended and adjust density to 5 * 10 with the mesenchymal stem cell serum-free culture medium of above-mentioned preparation
6Cells/ml is inoculated in 6 orifice plates, puts 37 ℃ of 5%C0
2Cultivate in the saturated humidity incubator, remove the mesenchymal stem cell serum-free culture medium of the fresh above-mentioned preparation of supernatant liquor replacing every three days and adjust density to 5 * 10
6Cells/ml cultivates after 10 days cell degree of converging and reaches 85% o'clock digestion harvested cell.
UCMSCs (umbilical cord mesenchymal stem cells) vitro culture: after the full-term pregnancy childbirth fetal cord of obtaining aseptic fully washs the stain of dehematizing, reject the umbilical cord arteriovenous, dissect to scrape and get Wharton ' s Jelly (stroma), shred into about 1mm
3The tissue block of size, normal saline flushing, resuspended with the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation, be inoculated in the culturing bottle, cell degree of converging reaches 85% o'clock digestion harvested cell after 14 days.
Distinguish resuspended with the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation BMSCs and the UCMSCs of above-mentioned vitro culture and adjustment density 1 * 10
6Cells/ml is inoculated in respectively in the culturing bottle, and cell degree of converging reaches 85% o'clock digestion harvested cell after 3 days, for the cell cycle detect and colony to form detection standby.
Cell cycle is detected:
Above-mentioned standby BMSCs and UCMSCs cultivated for 2 generations in the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation after, with 0.5 * 10
4The density of cells/mL is inoculated in the 100mm culture dish, cultivate harvested cell behind the 6d respectively, 70% ethanol with-20 ℃ of precoolings is fixed, the dyeing of 50 μ g/mL iodate third ingots (PI), flow cytometer (BD company) detects 10000 cells, the result is as scheming: the mesenchymal stem cells MSCs G0/G1 phase 48.9%, the G2/M phase 14.5%, the S phase 36.6%, the umbilical cord mesenchymal stem cells G0/G1 phase 49.8%, the G2/M phase 15.8%, the S phase 34.4%, a high proportion of G0/G1 phase cell shows that the mescenchymal stem cell in these two kinds of sources has the differentiation potential of height.
Colony forms and detects:
Above-mentioned standby BMSCs and UCMSCs cultivated for 2 generations in the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation after, density with 200cells/well is inoculated in 6 orifice plates that overlay 0.2% gelatin, every 3d changes liquid, the purple dyeing of 7d post crystallization colony, meter colony number under the phase microscope, surpass 50 cells and count a colony, every group establish 3 parallel.
Calculate colony forming efficiency by following formula:
Colony forming efficiency (Colony-forming efficiency, CFE) (%)=(colony number/inoculating cell number) * 100%
The result shows: the marrow BMSCs after the serum-free amplification and the colony forming efficiency of umbilical cord UCMSCs are respectively (13.67% ± 4.04%) and (15.00% ± 2.00%), and the colony of cell forms the ability no significant difference.Deng oozing the violet staining cell colony, the colony size of seeing two groups of cells is no significant difference also.
The preparation of 1L mesenchymal stem cell serum-free perfect medium: with product type be Gibco 884400 DMEM/F12 dry powder (packing: 10 bags of * 1L/ boxes) 1 bag, recombinant human insulin 2mg, human serum albumin 30g, human transferrin 10mg, cholesterol 10mg, Sodium Selenite 0.05mg, iron nitrate 10mg, dextran 10mg mixes, and adds injection water 800ml, stirring and dissolving, and regulate pH to 7.1 ± 0.1 with the hydrochloric acid of 0.5mol/L, be settled to 1L, with the degerming of 0.22um membrane filtration, deposit for 4 ℃ behind the mixing.
BMSCs (mesenchymal stem cells MSCs) vitro culture: extract volunteer's marrow 10ml by hospital, equal-volume is laid on the lymphocyte separation medium of 1.073g/ml, and the centrifugal 30min of 2250rpm gets cloud mononuclearcell layer, washes 3 times with physiological saline; Again successively with 2000rpm, 1800rpm, the centrifugal supernatant liquor that goes of 1600rpm, each five minutes, that the cell that goes to obtain after the supernatant liquor is resuspended and adjust density to 5 * 10 with the mesenchymal stem cell serum-free culture medium of above-mentioned preparation
6Cells/ml is inoculated in 6 orifice plates, puts in 37 ℃ of 5%CO2 saturated humidity incubators and cultivates, and removes the mesenchymal stem cell serum-free culture medium of the fresh above-mentioned preparation of supernatant liquor replacing every three days and adjusts density to 5 * 10
6Cells/ml cultivates after 10 days cell degree of converging and reaches 85% o'clock digestion harvested cell.
UCMSCs (umbilical cord mesenchymal stem cells) vitro culture: after the full-term pregnancy childbirth fetal cord of obtaining aseptic fully washs the stain of dehematizing, reject the umbilical cord arteriovenous, dissect to scrape and get Wharton ' s Jelly (stroma), shred into about 1mm
3The tissue block of size, normal saline flushing, resuspended with the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation, be inoculated in the culturing bottle, cell degree of converging reaches 85% o'clock digestion harvested cell after 14 days.
Distinguish resuspended with the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation BMSCs and the UCMSCs of above-mentioned vitro culture and adjustment density 1 * 10
6Cells/m1 is inoculated in respectively in the culturing bottle, and cell degree of converging reaches 85% o'clock digestion harvested cell after 3 days, for the cell cycle detect and colony to form detection standby.
Cell cycle is detected:
Above-mentioned standby BMSCs and UCMSCs cultivated for 2 generations in the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation after, with 0.5 * 10
4The density of cells/mL is inoculated in the 100mm culture dish, cultivate harvested cell behind the 6d respectively, 70% ethanol with-20 ℃ of precoolings is fixed, the dyeing of 50 μ g/mL iodate third ingots (PI), flow cytometer (BD company) detects 10000 cells, the result is as scheming: the mesenchymal stem cells MSCs G0/G1 phase 44.6%, the G2/M phase 16.4%, the S phase 38.9%, the umbilical cord mesenchymal stem cells G0/G1 phase 36.4%, the G2/M phase 14.0%, the S phase 49.6%, a high proportion of G0/G1 phase cell shows that the mescenchymal stem cell in these two kinds of sources has the differentiation potential of height.
Colony forms and detects:
Above-mentioned standby BMSCs and UCMSCs cultivated for 2 generations in the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation after, density with 200cells/well is inoculated in 6 orifice plates that overlay 0.2% gelatin, every 3d changes liquid, the purple dyeing of 7d post crystallization colony, meter colony number under the phase microscope, surpass 50 cells and count a colony, every group establish 3 parallel.
Calculate colony forming efficiency by following formula:
Colony forming efficiency (Colony-forming efficiency, CFE) (%)=(colony number/inoculating cell number) * 100%
The result shows: the marrow BMSCs after the serum-free amplification and the colony forming efficiency of umbilical cord UCMSCs are respectively (17.67% ± 2.04%) and (16.00% ± 2.00%), and the colony of cell forms the ability no significant difference.Deng oozing the violet staining cell colony, the colony size of seeing two groups of cells is no significant difference also.
Embodiment 3
The preparation of 1L mesenchymal stem cell serum-free perfect medium: with product type be 12400-024 DMEM/F12 dry powder (packing: 10 bags of * 1L/ boxes) 1 bag, recombinant human insulin 10mg, human serum albumin 1g, human transferrin 1mg, cholesterol 50mg, Sodium Selenite 0.2mg, iron nitrate 1mg, dextran 100mg mixes, and adds injection water 800ml, stirring and dissolving, and regulate pH to 7.1 ± 0.1 with the hydrochloric acid of 0.5mol/L, be settled to 1L, with the degerming of 0.22um membrane filtration, deposit for 4 ℃ behind the mixing.
BMSCs (mesenchymal stem cells MSCs) vitro culture: extract volunteer's marrow 10ml by hospital, equal-volume is laid on the lymphocyte separation medium of 1.073g/ml, the centrifugal 30min of 2250rpm, get cloud mononuclearcell layer, wash 3 times with physiological saline, again successively with 2000rpm, 1800rpm, the centrifugal supernatant liquor that goes of 1600rpm, each five minutes; The cell that removes supernatant liquor is resuspended and adjust density to 5 * 10 with the mesenchymal stem cell serum-free culture medium of above-mentioned preparation
6Cells/ml is inoculated in 6 orifice plates, puts in 37 ℃ of 5%C02 saturated humidity incubators and cultivates, and removes the mesenchymal stem cell serum-free culture medium of the fresh above-mentioned preparation of supernatant liquor replacing every three days and adjusts density to 5 * 10
6Cells/ml cultivates after 10 days cell degree of converging and reaches 85% o'clock digestion harvested cell.
UCMSCs (umbilical cord mesenchymal stem cells) vitro culture: after the full-term pregnancy childbirth fetal cord of obtaining aseptic fully washs the stain of dehematizing, reject the umbilical cord arteriovenous, dissect to scrape and get Wharton ' s Jelly (stroma), shred into about 1mm
3The tissue block of size, normal saline flushing, resuspended with the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation, be inoculated in the culturing bottle, cell degree of converging reaches 85% o'clock digestion harvested cell after 14 days.
Distinguish resuspended with the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation BMSCs and the UCMSCs of above-mentioned vitro culture and adjustment density 1 * 10
6Cells/ml is inoculated in respectively in the culturing bottle, and cell degree of converging reaches 85% o'clock digestion harvested cell after 3 days, for the cell cycle detect and colony to form detection standby.
Cell cycle is detected:
Above-mentioned standby BMSCs and UCMSCs cultivated for 2 generations in the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation after, with 0.5 * 10
4The density of cells/mL is inoculated in the 100mm culture dish, cultivate harvested cell behind the 6d respectively, 70% ethanol with-20 ℃ of precoolings is fixed, the dyeing of 50 μ g/mL iodate third ingots (PI), flow cytometer (BD company) detects 10000 cells, the result is as scheming: the mesenchymal stem cells MSCs G0/G1 phase 45.4%, the G2/M phase 7.58%, the S phase 46.9%, the umbilical cord mesenchymal stem cells G0/G1 phase 44.9%, the G2/M phase 19.6%, the S phase 35.5%, a high proportion of G0/G1 phase cell shows that the mescenchymal stem cell in these two kinds of sources has the differentiation potential of height.
Colony forms and detects:
Above-mentioned standby BMSCs and UCMSCs cultivated for 2 generations in the mesenchymal stem cell serum-free perfect medium of above-mentioned preparation after, density with 200cells/well is inoculated in 6 orifice plates that overlay 0.2% gelatin, every 3d changes liquid, the purple dyeing of 7d post crystallization colony, meter colony number under the phase microscope, surpass 50 cells and count a colony, every group establish 3 parallel.
Calculate colony forming efficiency by following formula:
Colony forming efficiency (Colony-forming efficiency, CFE) (%)=(colony number/inoculating cell number) * 100%
The result shows: the marrow BMSCs after the serum-free amplification and the colony forming efficiency of umbilical cord UCMSCs are respectively (14.00% ± 1.64%) and (11.67% ± 3.00%), and the colony of cell forms the ability no significant difference.Deng oozing the violet staining cell colony, the colony size of seeing two groups of cells is no significant difference also.
Claims (3)
1. the serum-free perfect medium of a mescenchymal stem cell, constituted by basic medium and interpolation composition, it is characterized in that: described basic medium is DMEM/F12, described interpolation composition is by the recombinant human insulin, human serum albumin, human transferrin, cholesterol, Sodium Selenite, iron nitrate and dextran are formed, recombinant human insulin 2～10mg/L in the described interpolation composition, human serum albumin 1～30g/L, human transferrin 1～10mg/L, cholesterol 10～50mg/L, Sodium Selenite 0.05～0.2mg/L, iron nitrate 1～10mg/L, dextran 10～100mg/L, the pH of the serum-free perfect medium of described mescenchymal stem cell is 7～7.2.
2. the serum-free perfect medium of a kind of mescenchymal stem cell according to claim 1, it is characterized in that: the product type of described basic medium DMEM/F12 is 12400-024.
3. the serum-free perfect medium of a kind of mescenchymal stem cell according to claim 1 and 2, it is characterized in that: described serum-free perfect medium is used for the vitro culture mescenchymal stem cell, and described mescenchymal stem cell comprises mesenchymal stem cells MSCs, umbilical cord mesenchymal stem cells.
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| CN109182262A (en) * | 2018-09-25 | 2019-01-11 | 深圳市五零生命科技有限公司 | A kind of mesenchymal stem cell serum-free culture medium |
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| CN103243071B (en) * | 2013-05-09 | 2015-04-29 | 青岛瑞思德生物科技有限公司 | Clinical-grade human mesenchymal stem cell serum-free complete medium |
| CN103898043B (en) * | 2014-03-11 | 2016-05-11 | 山东大学附属千佛山医院 | Cell feeder layer and the application in cultivation people primary tumor cell thereof |
| CN104232573B (en) * | 2014-09-11 | 2017-01-11 | 安沂华 | Growth medium and use thereof, and method for cultivating umbilical cord mesenchymal stem cells |
| CN104694470B (en) * | 2015-03-31 | 2019-09-17 | 江苏普瑞康生物医药科技有限公司 | A kind of stem cell serum-free culture medium |
| CN105112365A (en) * | 2015-08-18 | 2015-12-02 | 广州暨南生物医药研究开发基地有限公司 | Serum-free medium for human umbilical cord mesenchymal stem cells and preparation method thereof |
| CN106754677A (en) * | 2016-12-24 | 2017-05-31 | 严志海 | A kind of external mesenchymal stem cells MSCs culture medium |
| CN106947736A (en) * | 2017-04-14 | 2017-07-14 | 青岛青春派生物科技有限公司 | A kind of human adipose mesenchymal stem cells Serum-free complete medium |
| CN106916784A (en) * | 2017-05-04 | 2017-07-04 | 刘力伟 | A kind of serum free medium and its application for human mesenchymal stem cell |
| CN108315296A (en) * | 2018-02-23 | 2018-07-24 | 深圳至博生物科技有限公司 | It the isolated culture method of mescenchymal stem cell and freezes, method for resuscitation |
| CN110343660A (en) * | 2018-04-08 | 2019-10-18 | 生物角（厦门）科技有限公司 | A kind of mesenchymal stem cell serum-free culture medium composition |
| CN108676773A (en) * | 2018-05-17 | 2018-10-19 | 广东芙金干细胞再生医学有限公司 | A kind of culture solution and preparation method delaying mescenchymal stem cell aging |
| CN110938590B (en) * | 2019-12-25 | 2021-02-23 | 依科赛生物科技（太仓）有限公司 | Mesenchymal stem cell serum-free medium and application thereof |
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| Title |
|---|
| 吴伟等.骨髓间充质干细胞无血清培养.《生物工程学报》.2009,第25卷(第1期), |
| 骨髓间充质干细胞无血清培养;吴伟等;《生物工程学报》;20090125;第25卷(第1期);121-128 * |
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| CN109182262A (en) * | 2018-09-25 | 2019-01-11 | 深圳市五零生命科技有限公司 | A kind of mesenchymal stem cell serum-free culture medium |
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