CN108517002A - A kind of stabilizer of stem cell culture supernatant secretory protein - Google Patents
A kind of stabilizer of stem cell culture supernatant secretory protein Download PDFInfo
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- CN108517002A CN108517002A CN201810360163.9A CN201810360163A CN108517002A CN 108517002 A CN108517002 A CN 108517002A CN 201810360163 A CN201810360163 A CN 201810360163A CN 108517002 A CN108517002 A CN 108517002A
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- stem cell
- cell culture
- culture supernatant
- stabilizer
- secretory protein
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
Abstract
The invention discloses a kind of stabilizers of stem cell culture supernatant secretory protein, including following raw material：4.8 8.7ml of glycerine, 6 6.9ug of vitamin C, 0.75 1g of xanthans, 0.3 0.68g of polyglyceryl fatty acid ester, 0.45 0.6g of carragheen, 0.5 0.86g of trehalose, 0.06 0.18g of mannitol, 1.2 1.8g of 0.4 0.7g of amino acid, 0.01 0.013g of high molecular cellulose and sorbierite.Raw material sources of the present invention are extensive, and finished product of the invention can keep the Activity and stabill of protein complex at normal temperatures, effectively solve the problems such as low temperature storage limits, freeze thawing and pot-life are short, meet the use demand of people.
Description
Technical field
The present invention relates to stem cell culture secretory protein field, specifically a kind of stem cell culture supernatant secretory protein it is steady
Determine agent.
Background technology
Stem cell is a kind of multipotential cell with the of self-replication capacity, and under certain condition, it can be divided into more
Kind functioning cell.Stage of development residing for stem cell is divided into embryonic stem cell and adult stem cell.According to the hair of stem cell
Potential is educated to be divided into three classes：Myeloid-lymphoid stem cell, multipotential stem cell and unipotent stem cell.Stem cell is a kind of inabundant differentiation, still not
There is ripe cell the potential function for regenerating various histoorgans and human body, medical field to be known as " general-purpose cell ".
Stem cell is widely used in medicine, the fields such as beauty research, as human umbilical cord mesenchymal stem cells are treated in recent years
The clinical research of diabetes, fat mesenchymal stem cell is for beauty and shaping field etc., and separately some researches show that stem cell is in life
Multiple protein, micromolecule polypeptide and growth regulator are secreted in growth process, long-time service can promote skin metabolism, resist
Wrinkle and anti-light aging, effect are more apparent.But since it is present in liquid, but -20 DEG C must be stored in, the pot-life is shorter, and
It is unable to multigelation.
The method that the preservation generally use freeze-dried powder low temperature of protein stores for a long time, keeps the activity and stabilization of protein
Property, that is, to ensure the stability of protein steric structure, the factor for influencing its stability usually has the pH value of solution, salinity
With the concentration of sugar, the effect of microorganism and the oxidation of air etc..On the one hand, amino acid is good antioxidant；Various sugar
It is active to play the role of protected protein matter；Ion chelating agent has multi-efficiency, such as anti-oxidant, can also prevent some ions heavy
Product；Glycerine, sorbierite can keep moistening for protein surface, it is made to be unlikely to inactivate because of dehydration.Other is water-soluble
The polymer (such as PEG) of foreign protein or chemically inert polymer substance, can be on antigen-antibody surface because its molecular weight is big
A kind of protective film is formed, makes protein structure from destroying, but these are all that could realize at low temperature, and pot-life
It is short.
Invention content
The purpose of the present invention is to provide a kind of stabilizers of stem cell culture supernatant secretory protein, to solve above-mentioned background
The problem of being proposed in technology.
To achieve the above object, the present invention provides the following technical solutions：
A kind of stabilizer of stem cell culture supernatant secretory protein, including following raw material：Glycerine 4.8-8.7ml, vitamin
C6-6.9ug, xanthans 0.75-1g, polyglyceryl fatty acid ester 0.3-0.68g, carragheen 0.45-0.6g, trehalose 0.5-
0.86g, mannitol 0.06-0.18g, amino acid 0.4-0.7g, high molecular cellulose 0.01-0.013g and sorbierite 1.2-
1.8g。
As a further solution of the present invention：Glycerine uses medical grade glycerine, vitamin C, xanthans, high molecular cellulose
And sorbierite is all made of sterile product.
As a further solution of the present invention：Glycerine, vitamin C, xanthans, high molecular cellulose and sorbierite it is pure
Degree is not less than 98%.
As a further solution of the present invention：Amino acid includes lysine, serine, aspartic acid, arginine and network ammonia
At least one of acid.
As a further solution of the present invention：Carragheen uses K- types or I- type carragheens, mannitol to use in kelp
The mannitol of extraction.
Compared with prior art, the beneficial effects of the invention are as follows：Raw material sources of the present invention are extensive, and finished product of the invention can be with
The Activity and stabill of protein complex at normal temperatures is kept, it is short effectively to solve low temperature storage limitation, freeze thawing and pot-life
The problems such as, meet the use demand of people.
Description of the drawings
Fig. 1 is the PH measurement charts when stabilizer of stem cell culture supernatant secretory protein uses.
Fig. 2 is the osmolarity figure when stabilizer of stem cell culture supernatant secretory protein uses.
Fig. 3 is the degradation rate figure when stabilizer of stem cell culture supernatant secretory protein uses.
Specific implementation mode
The technical solution of this patent is described in more detail With reference to embodiment.
Embodiment 1
A kind of stabilizer of stem cell culture supernatant secretory protein, including following raw material：Glycerine 4.8ml, vitamin C
6ug, xanthans 0.75g, polyglyceryl fatty acid ester 0.3g, carragheen 0.45g, trehalose 0.5g, mannitol 0.06g, amino acid
0.4g, high molecular cellulose 0.01g and sorbierite 1.2g.Glycerine uses medical grade glycerine, vitamin C, xanthans, macromolecule
Cellulose and sorbierite are all made of sterile product.Amino acid includes lysine, aspartic acid and arginic mixture.
Stem cell culture supernatant albumen composition is divided into two groups, experimental group adds the product of embodiment 1, and control group does not add
The product for adding embodiment 1, makes an addition to stabilizer normal saline, aseptic subpackaged at each 30,5ml/ branch, under normal temperature state
(20-25 DEG C) is sealed, its PH of periodic detection.The PH curve graphs of measurement are shown in Fig. 1.
Fig. 1 shows the albumen complex liquid for adding 1 product of embodiment under room temperature (20-25 DEG C), and fluctuation range is smaller, hence it is evident that
Better than the control group for not adding 1 product of embodiment.
Embodiment 2
A kind of stabilizer of stem cell culture supernatant secretory protein, including following raw material：Glycerine 6.6ml, vitamin C
6.5ug, xanthans 0.88g, polyglyceryl fatty acid ester 0.49g, carragheen 0.54g, trehalose 0.75g, mannitol 0.12g, ammonia
Base acid 0.55g, high molecular cellulose 0.012g and sorbierite 1.6g.Glycerine, vitamin C, xanthans, high molecular cellulose with
And the purity of sorbierite is not less than 98%.Carragheen uses K- type carragheens, mannitol to use the mannitol extracted in kelp.
Stem cell culture supernatant albumen composition is divided into two groups, experimental group adds the product of embodiment 2, and control group does not add
Add the product of embodiment 2, addition and stabilizer normal saline are aseptic subpackaged at each 30,5ml/ branch, under normal temperature state
(20-25 DEG C) is sealed, its osmolarity of periodic detection.The osmolarity curve graph of measurement is shown in Fig. 2.
Fig. 2 shows to add the product of embodiment 2 to osmolarity that two groups of results are in normal fluctuation range without influence
It is interior.
Embodiment 3
A kind of stabilizer of stem cell culture supernatant secretory protein, including following raw material：Glycerine 8.7ml, vitamin C
6.9ug, xanthans 1g, polyglyceryl fatty acid ester 0.68g, carragheen 0.6g, trehalose 0.86g, mannitol 0.18g, amino acid
0.7g, high molecular cellulose 0.013g and sorbierite 1.8g.Glycerine uses medical grade glycerine, vitamin C, xanthans, high score
Subbundle element and sorbierite are all made of sterile product.Glycerine, vitamin C, xanthans, high molecular cellulose and sorbierite
Purity is not less than 98%.Carragheen uses I- type carragheens, mannitol to use the mannitol extracted in kelp.Amino acid includes
The mixture of lysine, serine, aspartic acid, arginine and tyrosine.
Umbilical cord mesenchymal stem cells supernatant secretory protein liquid is divided into two groups, the product of one group of addition embodiment 3, one group is
Blank control, every group aseptic subpackaged 30,1.0ml/ branch is placed under room temperature, its total protein content of periodic detection.Total protein degradation
Rate diagram is shown in Fig. 3.
Fig. 3 shows that the umbilical cord mesenchymal stem cells supernatant for adding 3 product of embodiment can be obviously improved the degradation of albumen
Rate.
It is obvious to a person skilled in the art that invention is not limited to the details of the above exemplary embodiments, Er Qie
In the case of without departing substantially from spirit or essential attributes of the invention, the present invention can be realized in other specific forms.Therefore, no matter
From the point of view of which point, the present embodiments are to be considered as illustrative and not restrictive, and the scope of the present invention is by appended power
Profit requires rather than above description limits, it is intended that all by what is fallen within the meaning and scope of the equivalent requirements of the claims
Variation is included within the present invention.Any reference signs in the claims should not be construed as limiting the involved claims.
In addition, it should be understood that although this specification is described in terms of embodiments, but not each embodiment is only wrapped
Containing an independent technical solution, this description of the specification is merely for the sake of clarity, and those skilled in the art should
It considers the specification as a whole, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art
The other embodiment being appreciated that.
Claims (5)
1. a kind of stabilizer of stem cell culture supernatant secretory protein, which is characterized in that including following raw material：Glycerine 4.8-
8.7ml, vitamin C 6-6.9ug, xanthans 0.75-1g, polyglyceryl fatty acid ester 0.3-0.68g, carragheen 0.45-0.6g,
Trehalose 0.5-0.86g, mannitol 0.06-0.18g, amino acid 0.4-0.7g, high molecular cellulose 0.01-0.013g and mountain
Pears alcohol 1.2-1.8g.
2. the stabilizer of stem cell culture supernatant secretory protein according to claim 1, which is characterized in that the glycerine is adopted
With medical grade glycerine, vitamin C, xanthans, high molecular cellulose and sorbierite are all made of sterile product.
3. the stabilizer of stem cell culture supernatant secretory protein according to claim 1 or 2, which is characterized in that described sweet
Oil, vitamin C, xanthans, high molecular cellulose and sorbierite purity be not less than 98%.
4. the stabilizer of stem cell culture supernatant secretory protein according to claim 1, which is characterized in that the amino acid
Including at least one of lysine, serine, aspartic acid, arginine and tyrosine.
5. the stabilizer of stem cell culture supernatant secretory protein according to claim 1, which is characterized in that the carragheen
Using K- types or I- type carragheens, mannitol uses the mannitol extracted in kelp.
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| CN201810360163.9A CN108517002A (en) | 2018-04-20 | 2018-04-20 | A kind of stabilizer of stem cell culture supernatant secretory protein |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113403272A (en) * | 2021-07-23 | 2021-09-17 | 廊坊康宝汇泰生物技术有限公司 | Culture medium of primary umbilical cord mesenchymal stem cells and application thereof |
| CN114269359A (en) * | 2019-04-09 | 2022-04-01 | 康巴乔有限公司 | Methods for making and using mesenchymal stem cell-derived secreted proteomes |
| CN115399312A (en) * | 2022-09-02 | 2022-11-29 | 王嘉祥 | Preparation method of exosome normal-temperature storage protective agent in mesenchymal stem cell supernatant |
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| CN114269359A (en) * | 2019-04-09 | 2022-04-01 | 康巴乔有限公司 | Methods for making and using mesenchymal stem cell-derived secreted proteomes |
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| CN115399312A (en) * | 2022-09-02 | 2022-11-29 | 王嘉祥 | Preparation method of exosome normal-temperature storage protective agent in mesenchymal stem cell supernatant |
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