CN109439621A - A kind of mild enzyme cell dissociation buffer of stem cell - Google Patents
A kind of mild enzyme cell dissociation buffer of stem cell Download PDFInfo
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- CN109439621A CN109439621A CN201811340104.1A CN201811340104A CN109439621A CN 109439621 A CN109439621 A CN 109439621A CN 201811340104 A CN201811340104 A CN 201811340104A CN 109439621 A CN109439621 A CN 109439621A
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- 210000000130 stem cell Anatomy 0.000 title claims abstract description 34
- 210000004027 cell Anatomy 0.000 title claims abstract description 31
- 238000010494 dissociation reaction Methods 0.000 title claims abstract description 17
- 230000005593 dissociations Effects 0.000 title claims abstract description 17
- 239000000872 buffer Substances 0.000 title claims abstract description 12
- 108090000790 Enzymes Proteins 0.000 title claims description 12
- 102000004190 Enzymes Human genes 0.000 title claims description 12
- 230000001079 digestive effect Effects 0.000 claims abstract description 40
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 25
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims abstract description 18
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims abstract description 18
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 18
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 9
- 239000008103 glucose Substances 0.000 claims abstract description 9
- 239000001103 potassium chloride Substances 0.000 claims abstract description 9
- 235000011164 potassium chloride Nutrition 0.000 claims abstract description 9
- 235000017557 sodium bicarbonate Nutrition 0.000 claims abstract description 9
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims abstract description 9
- 239000011780 sodium chloride Substances 0.000 claims abstract description 9
- UEUXEKPTXMALOB-UHFFFAOYSA-J tetrasodium;2-[2-[bis(carboxylatomethyl)amino]ethyl-(carboxylatomethyl)amino]acetate Chemical compound [Na+].[Na+].[Na+].[Na+].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O UEUXEKPTXMALOB-UHFFFAOYSA-J 0.000 claims abstract description 9
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims abstract description 8
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical class [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims abstract description 6
- 238000004090 dissolution Methods 0.000 claims description 10
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 6
- 239000001257 hydrogen Substances 0.000 claims description 4
- 229910052739 hydrogen Inorganic materials 0.000 claims description 4
- DJFBJKSMACBYBD-UHFFFAOYSA-N phosphane;hydrate Chemical compound O.P DJFBJKSMACBYBD-UHFFFAOYSA-N 0.000 claims 3
- -1 disodium hydrogen Chemical class 0.000 claims 2
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical compound [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 claims 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims 1
- 108010019160 Pancreatin Proteins 0.000 abstract description 17
- 229940055695 pancreatin Drugs 0.000 abstract description 17
- 230000029087 digestion Effects 0.000 abstract description 15
- 238000000034 method Methods 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 4
- 238000004321 preservation Methods 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 2
- 230000003833 cell viability Effects 0.000 abstract 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 10
- 229940088598 enzyme Drugs 0.000 description 10
- 241001465754 Metazoa Species 0.000 description 7
- 239000004615 ingredient Substances 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 5
- 229910052757 nitrogen Inorganic materials 0.000 description 5
- 238000001914 filtration Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 3
- 229960000074 biopharmaceutical Drugs 0.000 description 3
- 238000001976 enzyme digestion Methods 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 210000000496 pancreas Anatomy 0.000 description 3
- 230000006798 recombination Effects 0.000 description 3
- 238000005215 recombination Methods 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 108090000317 Chymotrypsin Proteins 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 102000018690 Trypsinogen Human genes 0.000 description 1
- 108010027252 Trypsinogen Proteins 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000009087 cell motility Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229960002376 chymotrypsin Drugs 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- KCIDZIIHRGYJAE-YGFYJFDDSA-L dipotassium;[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate Chemical compound [K+].[K+].OC[C@H]1O[C@H](OP([O-])([O-])=O)[C@H](O)[C@@H](O)[C@H]1O KCIDZIIHRGYJAE-YGFYJFDDSA-L 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 238000003307 slaughter Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
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- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Developmental Biology & Embryology (AREA)
- Microbiology (AREA)
- Rheumatology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention discloses a kind of stem cell bland cell digestive ferment cell dissociation buffers, it is to be grouped as by the group of following concentration, each concentration unit is mg/L: dipotassium hydrogen phosphate 50~70, potassium chloride 350~450, sodium bicarbonate 300~400, sodium chloride 7000~9000, seven water disodium hydrogen phosphates 80~100, glucose 1000~2000, EDTA-4Na 200~500, the mild digestive ferment 1~120 of stem cell, surplus is water.The mild digestive ferment cell dissociation buffer of stem cell of the invention is better than traditional pancreatin to the digestion power of cell, and general cell can digest completely within 3 minutes.Cell dissociation process is less to the damage of cell, inoculates after the completion of digestion, and Cell viability is high, growth conditions are good.The mild digestive ferment cell dissociation buffer of stem cell of the invention, without -20 DEG C of preservations, using effect is constant within saving 12 months in 2~8 DEG C.
Description
Technical field
The present invention relates to a kind of mild enzymic digestion liquid of stem cell, are exclusively used in stem cell digestion, belong to bio-pharmaceuticals, biology is controlled
The related fieldss such as treatment.
Background technique
Trypsase is a kind of proteolytic enzyme of animal origin, is activated and is produced by the trypsinogen of animal pancreatic secretion
Raw, the hydrolysis arginine and lysine of energy specificity complete the peptide bond of cardinal extremity, are widely used in field of biotechnology and medical treatment neck
Domain.
There are mainly two types of the approach for obtaining trypsase at present, and traditional mode is obtained from animal, is passed through
Slaughter ox, sheep, pig etc. extract trypsase from its pancreas.In addition, obtaining tryptose in the way of microbial fermentation
Enzyme gradually has been favored by people because efficient and specificity is strong.
With the continuous development of bio-pharmaceuticals, trypsase becomes the main raw and auxiliary material of some bio-pharmaceuticals.Most because of it
Finished product will be used for human body diseases treatment, and the quality of quality may will have a direct impact on the life and health of the mankind.Due to pancreas egg
White enzyme source is easy in organism, complicated component, donor by pathogen contamination, once this pollution is not thoroughly removed
And it is brought into the production of pancreatin, it is possible to will lead to serious consequence.
Currently, yeast and Escherichia coli are the synthesis common microbial hosts of trypsase.Recombinant trypsin is to utilize
The trypsase that recombinant DNA technology goes out in expression of recombinant e. coli, without any animal derived component, and background understands, purity
It is higher, thus it is suitable for free serum culture, it can be applied to the production of antibody and drug.
Summary of the invention
For the above-mentioned prior art, the present invention provides a kind of mild enzymic digestion liquid of stem cell, it is an object of the invention to
Digestion operation is carried out to stem cells free serum culture, to stem cell action temperature and, it is not damaged, while avoiding conventional animal source
The intervention of trypsase, to guarantee that entire cell cultivation process does not have participating in for animal sources component.Meanwhile this product can be 2~8
It is saved for a long time in DEG C environment, it is used as needed, it is intended to bring bigger convenience for experimental implementation person.
The present invention is achieved by the following technical solutions:
A kind of mild digestive ferment digestive juice of stem cell, is grouped as by the group of following concentration, and each concentration unit is mg/L: phosphoric acid
Hydrogen dipotassium 50~70, potassium chloride 350~450, sodium bicarbonate 300~400, sodium chloride 7000~9000, seven water disodium hydrogen phosphates
80~100, glucose 1000~2000, EDTA-4Na 200~500, the mild digestive ferment 1~120 of stem cell, surplus is water.
The mild digestive ferment is purchased from KERRY, name of product: rTrypsin, product article No.: 30624799.5GR.
The mild digestive ferment, is produced by expression of recombinant e. coli, the cation of amino acid sequence and human pancreas source
Type trypsase is completely the same, and digestion specificity is identical；It, can not in no natural pancreatin of tradition without any animal-based components
The components such as the chymotrypsin avoided.
The mild digestive ferment digestive juice of stem cell of the invention the preparation method comprises the following steps: above-mentioned component in addition to water is taken, according to it
Respective dissolution characteristics classification dissolution, then mixes, be added water make each component final concentration as described above, adjust pH value to 7.2~
7.4 to get, industrial filter element filtering, while nitrogen protection is dispensed and (unstability ingredient avoided to be oxidized).
The mild digestive ferment cell dissociation buffer of stem cell of the invention is better than traditional pancreatin to the digestion power of cell, generally
It can be digested within cell 3 minutes completely.Cell dissociation process is less to the damage of cell, inoculates after the completion of digestion, cell
Motility rate is high, growth conditions are good.The mild digestive ferment cell dissociation buffer of stem cell of the invention, without -20 DEG C of preservations, at 2~8 DEG C
Using effect is constant within middle preservation 12 months.Preparation process of the invention all uses on-line cleaning and online sterilizing installation,
Guarantee the stabilization of difference between batch, while using inert body nitrogen protection in production, avoid unstability ingredient from being oxidized, the technique is raw
The immunocyte serum free medium quality of production is stablized, and solves the problems, such as that most of producer's qualities of production are not easy to control.
Detailed description of the invention
Fig. 1: traditional pancreatin liquid chromatogram.
Fig. 2: the mild digestive ferment liquid chromatogram of stem cell.
Before Fig. 3: MSC digestion.
After Fig. 4: MSC digestion.
Specific embodiment
Below with reference to embodiment, the present invention is further illustrated.
Instrument involved in following embodiments, reagent, material etc. are unless otherwise noted existing in the prior art
Conventional instrument, reagent, material etc., can be obtained by regular commercial sources.Experimental method involved in following embodiments, inspection
Survey method etc. is unless otherwise noted existing routine experiment method in the prior art, detection method etc..
Mild digestive ferment used in the present invention is purchased from KERRY, name of product: rTrypsin, product article No.:
30624799.5GR。
Embodiment 1 prepares the mild enzyme cell dissociation buffer of stem cell
It is to be grouped as by the group of following concentration, each concentration unit is mg/L: dipotassium hydrogen phosphate 60, potassium chloride 400, sodium bicarbonate
350, sodium chloride 8000, seven water disodium hydrogen phosphates 90, glucose 1500, EDTA-4Na 350, the mild digestive ferment 30 of stem cell,
Surplus is water.
The preparation method comprises the following steps: taking above-mentioned component in addition to water, according to its respectively dissolution characteristics classification dissolution, then mixes, add
Enter water and make each component final concentration as described above, adjust pH value to 7.2~7.4 to get, industrial filter element filtering, while nitrogen protection
It is dispensed and (unstability ingredient is avoided to be oxidized).
Embodiment 2 prepares the mild digestive ferment cell dissociation buffer of stem cell
It is to be grouped as by the group of following concentration, each concentration unit is mg/L: dipotassium hydrogen phosphate 50, potassium chloride 450, sodium bicarbonate
300, sodium chloride 9000, seven water disodium hydrogen phosphates 80, glucose 2000, EDTA-4Na 500, the mild digestive ferment 10 of stem cell,
Surplus is water.
The preparation method comprises the following steps: taking above-mentioned component in addition to water, according to its respectively dissolution characteristics classification dissolution, then mixes, add
Enter water and make each component final concentration as described above, adjust pH value to 7.2~7.4 to get, industrial filter element filtering, while nitrogen protection
It is dispensed and (unstability ingredient is avoided to be oxidized).
Embodiment 3 prepares the mild digestive ferment cell dissociation buffer of stem cell
It is to be grouped as by the group of following concentration, each concentration unit is mg/L: dipotassium hydrogen phosphate 70, potassium chloride 350, sodium bicarbonate
400, sodium chloride 7000, seven water disodium hydrogen phosphates 100, glucose 2000, EDTA-4Na 200, the mild digestive ferment of stem cell
50, surplus is water.
The preparation method comprises the following steps: taking above-mentioned component in addition to water, according to its respectively dissolution characteristics classification dissolution, then mixes, add
Enter water and make each component final concentration as described above, adjust pH value to 7.2~7.4 to get, industrial filter element filtering, while nitrogen protection
It is dispensed and (unstability ingredient is avoided to be oxidized).
Test the mild digestive ferment digestive juice vitellophag of stem cell
Using the mild digestive ferment vitellophag of stem cell of the invention, meanwhile, with traditional pancreatin (traditional pancreatin, refer to directly from
Obtained pancreatin is extracted in pig body) as control.
(1) stability contrast: the stability of the mild digestive ferment of stem cell used in the comparison present invention and traditional pancreatin, such as table
Shown in 1.
Table 1
(2) comparison or purity: as shown in Figure 1 and Figure 2.As seen from Figure 1, traditional pancreatin impurity is more (peak is more, and impurity is more), tradition
The higher ingredient of the visible four kinds of contents of pancreatin can not be separated, be degraded, surely simultaneously because one of impurity content is higher
Qualitative difference.From Figure 2 it can be seen that mild digestive ferment almost free from admixture, purity is high, peak type is good, and stability is good, no degradation peak.
(3) application method compares: as shown in table 2.
Table 2
(4) digestion time compares: comparing mild digestive ferment of the invention, traditional pancreatin and Gibco(recombination pancreatin) (commodity
Change product, market purchase) to the digestion time of cell, as shown in table 2.
Table 3
| Cell line title | The mild enzyme of the present invention | Gibco (recombination pancreatin) | Traditional pancreatin |
| MSC (mescenchymal stem cell) | 3min | 2min | 1min |
Seen from table 3, the recombination pancreatin used in the present invention, shortens the cell dissociation time, has to the listed cell dissociation time
It significantly improves, substantially reduces the triviality of work；While shortening the cell dissociation time, it is also ensured that postdigestive thin
Born of the same parents' motility rate is still greater than 90% or more.
Compare mild enzyme (different amounts) of the invention to the digestion time of cell, as shown in table 4.
Table 4
| Cell line title | Mild enzyme 10mg/L | Mild enzyme 60mg/L | Pancreatin 120mg/L |
| MSC (mescenchymal stem cell) | 3min | 1min | Excessively digestion |
By table 4 as it can be seen that the mild enzyme of different ratio is larger to impact cell, the usage amount through screening 10mg/L is optimal.
(5) mild enzymic digestion cell of the invention, comparison digestion front and back, as shown in figs. 34, by scheming digestion effect: are used
It is found that after mild enzymatic treatment, cell can normal stool, activity of pancreatic enzyme is obvious.
Above-mentioned, although specific embodiments of the present invention have been described in conjunction with the embodiments, not protects to the present invention
The limitation of range, those skilled in the art should understand that, based on the technical solutions of the present invention, those skilled in the art
The various modifications or changes that can be made are not needed to make the creative labor still within protection scope of the present invention.
Claims (6)
1. a kind of mild digestive ferment enzyme cell dissociation buffer of stem cell, it is characterised in that: be to be grouped as by the group of following concentration, respectively
Concentration unit is mg/L: dipotassium hydrogen phosphate 50~70, potassium chloride 350~450, sodium bicarbonate 300~400, sodium chloride 7000~
9000, seven water disodium hydrogen phosphates 80~100, glucose 1000~2000, EDTA-4Na 200~500, mild digestive ferment 1~
120, surplus is water.
2. the mild digestive ferment digestive juice of stem cell according to claim 1, it is characterised in that: be by the component of following concentration
Composition, each concentration unit is mg/L: dipotassium hydrogen phosphate 60, potassium chloride 400, sodium bicarbonate 350, sodium chloride 8000, seven water phosphorus
Sour disodium hydrogen 90, glucose 1500, EDTA-4Na 350, mild digestive ferment 10, surplus is water.
3. the mild digestive ferment digestive juice of stem cell according to claim 1, it is characterised in that: be by the component of following concentration
Composition, each concentration unit is mg/L: dipotassium hydrogen phosphate 50, potassium chloride 450, sodium bicarbonate 300, sodium chloride 9000, seven water phosphorus
Sour disodium hydrogen 80, glucose 2000, EDTA-4Na 500, mild digestive ferment 60, surplus is water.
4. the mild digestive ferment digestive juice of stem cell according to claim 1, it is characterised in that: be by the component of following concentration
Composition, each concentration unit is mg/L: dipotassium hydrogen phosphate 70, potassium chloride 350, sodium bicarbonate 400, sodium chloride 7000, seven water phosphorus
Sour disodium hydrogen 100, glucose 2000, EDTA-4Na 200, mild digestive ferment 120, surplus is water.
5. the preparation method of the mild digestive ferment digestive juice of stem cell according to any one of claims 1 to 4, it is characterised in that:
Component in addition to water is taken, according to its respectively dissolution characteristics classification dissolution, is then mixed, water, which is added, makes each component final concentration such as right
It is required that described in any one of 1~4, adjust pH value to 7.2~7.4 to get.
6. application of the mild digestive ferment cell dissociation buffer of stem cell according to any one of claims 1 to 4 in vitellophag.
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| CN201811340104.1A CN109439621A (en) | 2018-11-12 | 2018-11-12 | A kind of mild enzyme cell dissociation buffer of stem cell |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811340104.1A CN109439621A (en) | 2018-11-12 | 2018-11-12 | A kind of mild enzyme cell dissociation buffer of stem cell |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111690601A (en) * | 2020-06-08 | 2020-09-22 | 海南优尼科尔生物科技有限公司 | Preparation method of umbilical cord mesenchymal stem cell preparation |
| CN112680412A (en) * | 2021-01-14 | 2021-04-20 | 辽宁省肿瘤医院 | Animal source-free passage method for passage stem cells |
| CN113088486A (en) * | 2021-03-03 | 2021-07-09 | 广东为象生物科技有限公司 | Enzyme-free dissociation liquid for separating adherent cells and preparation method thereof |
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| US20060188892A1 (en) * | 2005-02-18 | 2006-08-24 | Ambion, Inc. | Enzymatic digestion of tissue |
| CN107090427A (en) * | 2017-06-23 | 2017-08-25 | 友康恒业生物科技（北京）有限公司 | A kind of genetic recombination trypsase cell dissociation buffer |
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2018
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US20060188892A1 (en) * | 2005-02-18 | 2006-08-24 | Ambion, Inc. | Enzymatic digestion of tissue |
| CN107090427A (en) * | 2017-06-23 | 2017-08-25 | 友康恒业生物科技（北京）有限公司 | A kind of genetic recombination trypsase cell dissociation buffer |
Non-Patent Citations (1)
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111690601A (en) * | 2020-06-08 | 2020-09-22 | 海南优尼科尔生物科技有限公司 | Preparation method of umbilical cord mesenchymal stem cell preparation |
| CN112680412A (en) * | 2021-01-14 | 2021-04-20 | 辽宁省肿瘤医院 | Animal source-free passage method for passage stem cells |
| CN112680412B (en) * | 2021-01-14 | 2024-02-23 | 辽宁省肿瘤医院 | Animal source-free passage method for passage of stem cells |
| CN113088486A (en) * | 2021-03-03 | 2021-07-09 | 广东为象生物科技有限公司 | Enzyme-free dissociation liquid for separating adherent cells and preparation method thereof |
| CN113088486B (en) * | 2021-03-03 | 2023-06-16 | 广东为象生物科技有限公司 | Non-enzymatic hydrolysate for separating adherent cells and preparation method thereof |
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