CN109010369A - A kind of preparation method for repairing mammalian gut damage freeze-dried powder - Google Patents
A kind of preparation method for repairing mammalian gut damage freeze-dried powder Download PDFInfo
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- CN109010369A CN109010369A CN201811127256.3A CN201811127256A CN109010369A CN 109010369 A CN109010369 A CN 109010369A CN 201811127256 A CN201811127256 A CN 201811127256A CN 109010369 A CN109010369 A CN 109010369A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- 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
- C12N5/0665—Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood
Abstract
The present invention provides a kind of preparation methods of reparation mammalian gut damage freeze-dried powder, it is related to biological stem cells technology field, pass through screening and culturing object and separate, umbilical cord cleaning and processing, umbilical cord tissue culture, mescenchymal stem cell secondary culture, the processing of mescenchymal stem cell culture supernatant and freeze-dried powder the multiple process means such as preparation, realize the reagent for preparing the cytokine profiles that the umbilical cord mesenchymal stem cells containing high activity for repairing mammalian gut damage are secreted.
Description
Technical field
The present invention relates to biological stem cells technology fields more particularly to a kind of mammalian gut of repairing to damage freeze-dried powder
Preparation method.
Background technique
Mescenchymal stem cell refers to a kind of stem cell subgroup with triploblastica differentiation potential, in certain circumstances, theoretical
On can be induced to differentiate into human body any one histocyte, be mainly derived from the interstitial tissue being widely present in human body, especially
Marrow, fat, umbilical cord and placenta etc., since it is with powerful proliferative capacity, stronger differentiation capability, low immunogenicity and
Immunoloregulation function, application value clinically are increasingly paid close attention to by people.It is studied not with to mescenchymal stem cell
Disconnected to go deep into, secretion factor has become the hot spot of research.Mescenchymal stem cell can secrete cytokine profiles, these cells because
Son can effectively regulate and control body cell signal transduction, activating human body stem cell, and then physiological reparation or substitution body injury, disease
The cell of change and aging.
Some researches show that umbilical cord mesenchymal stem cells can secrete cytokine profiles during culture, epidermis is such as secreted
Growth factor (Epidermal Growth Factor, EGF), hepatocyte growth factor (Hepatocyte Growth
Factor, HGF), tumor necrosis factor (Tumor Necrosis Factor, TNF), vascular endothelial growth factor
(Vascularendothelial Growth Factor, VEGF), nerve growth factor (Nerve Growth Factor,
NGF) etc., these cell factors can promote dermal cell growth to a certain extent, improve cell growth microenvironment, promote wound
Mouth healing delays senescence, improves the effects of skin growth state, but is typically prepared and damages for repairing mammalian gut
When universal activity decline, take that effect is unobvious, thus how to prepare for repair mammalian gut damage containing height
It is urgently to be resolved that the reagent of the cytokine profiles of active umbilical cord mesenchymal stem cells secretion becomes those skilled in the art
Problem.
Summary of the invention
The technical problem to be solved by the present invention is to overcome the deficiencies in the prior art, it is dynamic to provide a kind of reparation lactation
The preparation method of object intestinal tract injury freeze-dried powder, the preparation method are able to produce out for repairing containing for mammalian gut damage
The cytokine profiles freeze-dried powder of the umbilical cord mesenchymal stem cells secretion of high activity.
The present invention is to be achieved by the following technical programs: a kind of preparation for repairing mammalian gut damage freeze-dried powder
Method, the following steps are included:
(1) screening and culturing object and separate: the screening of umbilical cord is to choose pregnant woman age in 22-28 one full year of life, pregnancy period≤40 week,
The neonatal umbilical cord of health full term is placed in close to baby end about 20cm or so containing dual anti-umbilical cord acquisition liquid preservation, in ice chest
It transports as early as possible；
(2) it umbilical cord cleaning and processing: is placed in sterile cup ware with 75% alcohol Rapid Cleaning one time, then with containing dual anti-nothing
Bacterium physiological saline repeated flushing, until without obvious clot；Umbilical cord is longitudinally splitted, blood vessel is rejected；With containing dual anti-sterile physiological
Umbilical cord is divided segment to shred and is placed in sterile cup ware by salt water by tissue block rinsed clean with eye scissors, further by umbilical cord
It shreds to fine tissue block；
(3) umbilical cord tissue culture: being inoculated in culture bottle bottom according to suitable density for tissue block, adds suitable containing blood
The DMEM/F12 culture solution of clear sub, by body turnover bottle bottom think on be placed in 37 DEG C, it is adherent in the incubator of 5% CO2
4h afterwards turns over body bottom of bottle and is statically placed in incubator downwards and cultivates, routine observation replacement or addition culture solution；8d or so is aobvious
Micro- microscopic observation, it is seen that have into fibrous cell around tissue block and climb out of, removal tissue block replacement culture solution continues to cultivate the left side 2d
The right side, cell is P0 at this time, and attached cell is transferred in new culture bottle with TrypLE digestion and is cultivated, and cell is P1 at this time；
(4) mescenchymal stem cell secondary culture: the cell separated in step (3) is continued into secondary culture to P6, passage is
When cell growth reaches 80% fusion in culture bottle, TrypLE vitellophag, 1: 4 secondary culture starts to collect P2 to P6's
Culture supernatant；
(5) centrifugation of mescenchymal stem cell culture supernatant, purifying and filtration sterilization: will in (4) collection culture supernatant into
Row high speed centrifugation, again with supernatant after the centrifugation of sterile apyrogeneity container collection；By centrifuged supernatant concentration, then passed through
Device is filtered to carry out degerming to concentrate and dispense into sterile apyrogeneity bottle；
(6) preparation of freeze-dried powder: KFG and freeze drying protectant is added in frozen dried concentrate in concentrate liquid；Through cold
Mescenchymal stem cell factor freeze-dried powder is made after lyophilization processing.
According to the above technical scheme, it is preferable that using DMEM/F12 (HEPES, the NO PHENOL for containing 1% serum substitute
RED, L-GLUTAMINE) culture solution culture human umbilical cord mesenchymal stem cells.
According to the above technical scheme, it is preferable that the diameter dimension that the umbilical cord scissors in step (1) is broken to fine tissue block is
2mm-4mm。
According to the above technical scheme, it is preferable that the culture bottle of step (3) be T75cm2The surface sterile apyrogeneity TC is ventilative
Type culture bottle, the culture bottle for being passaged to P1 is T175cm2The surface sterile apyrogeneity TC vapor-permeable type culture bottle.
According to the above technical scheme, it is preferable that the culture supernatant collected in step (5) is centrifuged with 3000r/min
30min takes centrifuged supernatant, and the supernatant repeatedly collected is stored in -20 DEG C.
According to the above technical scheme, it is preferable that the culture supernatant filtering and concentrating of step (5) is the culture supernatant that will be collected
Liquid removes participation cell in thaw at RT, by 0.45um membrane filtration, by filter membrane ultrafiltration collect molecular weight 3~
The concentrate of the cell factor of 30KD is collected concentrate by 0.22um membrane filtration degerming.
According to the above technical scheme, it is preferable that it is 0.01% that KFG ratio is added in step (6)；Freeze drying protectant is at subpackage
It includes: trehalose, mannitol, dextran and human serum albumins.
According to the above technical scheme, it is preferable that after freeze drying protectant is added in step (6), trehalose is in concentrate
Percent by volume is 1~8%, and percent by volume of the mannitol in concentrate is 1~8%, and dextran is in concentrate
Percent by volume is 1~5%, and percent by volume of the human serum albumins in concentrate is 0.1~2%.
According to the above technical scheme, it is preferable that it is dry thin that mesenchyma is made after the chilled dry drying process in step (6)
Intracellular cytokine freeze-dried powder, storage temperature are 2 DEG C~4 DEG C.
The beneficial effects of the present invention are:
(1) present invention preferably source of the human umbilical cord mesenchymal stem cells as active cytokine, mescenchymal stem cell
In supernatant be rich in epidermal growth factor, vascular endothelial growth factor and stem cell factor isoreactivity factor component, the activity because
Son can promote the growth and proliferation of internal stem cell, and cell tissue in enteron aisle etc. is promoted to accelerate growth rate.By this hair
The cell factor stoste purity is high of bright purifying, protein content is high, and repairing effect is significant.
(2) present invention securely and reliably avoids traditional blood using the culture medium culture umbilical cord stem cells containing serum substitute
Clear cultivation may caused immunological rejection.
(3) dry powder is made in stem cell factor by the present invention, can be saved under cryogenic for a long time；Before use,
Deionized water and dry powder can be mixed and be used, solve the problems, such as that stem cell factor cannot save for a long time.
Detailed description of the invention
Fig. 1 shows the preparation flow schematic diagram of embodiment according to the present invention.
Specific embodiment
In order to make those skilled in the art more fully understand technical solution of the present invention, with reference to the accompanying drawing and most
The present invention is described in further detail for good embodiment.
As shown, the present invention provides a kind of preparation methods of reparation mammalian gut damage freeze-dried powder, including with
Lower step:
(1) screening and culturing object and separate: the screening of umbilical cord is to choose pregnant woman age in 22-28 one full year of life, pregnancy period≤40 week,
The neonatal umbilical cord of health full term is placed in close to baby end about 20cm or so containing dual anti-umbilical cord acquisition liquid preservation, in ice chest
It transports as early as possible；
(2) it umbilical cord cleaning and processing: is placed in sterile cup ware with 75% alcohol Rapid Cleaning one time, then with containing dual anti-nothing
Bacterium physiological saline repeated flushing, until without obvious clot；Umbilical cord is longitudinally splitted, blood vessel is rejected；With containing dual anti-sterile physiological
Umbilical cord is divided segment to shred and is placed in sterile cup ware by salt water by tissue block rinsed clean with eye scissors, further by umbilical cord
It shreds to fine tissue block；
(3) umbilical cord tissue culture: being inoculated in culture bottle bottom according to suitable density for tissue block, adds suitable containing blood
The DMEM/F12 culture solution of clear sub, by body turnover bottle bottom think on be placed in 37 DEG C, it is adherent in the incubator of 5% CO2
4h afterwards turns over body bottom of bottle and is statically placed in incubator downwards and cultivates, routine observation replacement or addition culture solution；8d or so is aobvious
Micro- microscopic observation, it is seen that have into fibrous cell around tissue block and climb out of, removal tissue block replacement culture solution continues to cultivate the left side 2d
The right side, cell is P0 at this time, and attached cell is transferred in new culture bottle with TrypLE digestion and is cultivated, and cell is P1 at this time；
(4) mescenchymal stem cell secondary culture: the cell separated in step (3) is continued into secondary culture to P6, passage is
When cell growth reaches 80% fusion in culture bottle, TrypLE vitellophag, 1: 4 secondary culture starts to collect P2 to P6's
Culture supernatant；
(5) centrifugation of mescenchymal stem cell culture supernatant, purifying and filtration sterilization: will in (4) collection culture supernatant into
Row high speed centrifugation, again with supernatant after the centrifugation of sterile apyrogeneity container collection；By centrifuged supernatant concentration, then passed through
Device is filtered to carry out degerming to concentrate and dispense into sterile apyrogeneity bottle；
(6) preparation of freeze-dried powder: KFG and freeze drying protectant is added in frozen dried concentrate in concentrate liquid；Through cold
Mescenchymal stem cell factor freeze-dried powder is made after lyophilization processing.
According to above-described embodiment, it is preferable that using DMEM/F12 (HEPES, the NO PHENOL for containing 1% serum substitute
RED, L-GLUTAMINE) culture solution culture human umbilical cord mesenchymal stem cells.
According to above-described embodiment, it is preferable that the diameter dimension that the umbilical cord scissors in step (1) is broken to fine tissue block is 2mm
~4mm.
According to above-described embodiment, it is preferable that the culture bottle of step (3) be T75cm2The surface sterile apyrogeneity TC vapor-permeable type
Culture bottle, the culture bottle for being passaged to P1 is T175cm2The surface sterile apyrogeneity TC vapor-permeable type culture bottle.
According to above-described embodiment, it is preferable that the culture supernatant collected in step (5) is to be centrifuged 30min with 3000r/min
Centrifuged supernatant is taken, the supernatant repeatedly collected is stored in -20 DEG C.
According to above-described embodiment, it is preferable that the culture supernatant filtering and concentrating of step (5) is the culture supernatant that will be collected
In thaw at RT, participation cell is removed by 0.45um membrane filtration, molecular weight is being collected in 3~30KD by filter membrane ultrafiltration
Cell factor concentrate, pass through 0.22um membrane filtration degerming, collect concentrate.
According to above-described embodiment, it is preferable that it is 0.01% that KFG ratio is added in step (6)；Freeze drying protectant is at subpackage
It includes: trehalose, mannitol, dextran and human serum albumins.
According to above-described embodiment, it is preferable that after freeze drying protectant is added in step (6), body of the trehalose in concentrate
Product percentage is 1~8%, and percent by volume of the mannitol in concentrate is 1~8%, body of the dextran in concentrate
Product percentage is 1~5%, and percent by volume of the human serum albumins in concentrate is 0.1~2%.
According to above-described embodiment, it is preferable that mescenchymal stem cell is made after the chilled dry drying process in step (6)
Factor freeze-dried powder, storage temperature are 2 DEG C -4 DEG C.
A kind of coherent detection for repairing mammalian gut damage freeze-dried powder:
The identification of umbilical cord mesenchymal stem cells: collecting P2 cell, with flow cytometry cell surface antigen, as a result table
Clear-cells can express cells and characteristic of stem surface antigen CD73, CD90, CD105 etc., fail express surface antigen CD19,
CD34, CD45 etc., it was demonstrated that the cell of culture is human mesenchymal stem cell.
The determining the protein quantity of human umbilical cord mesenchymal stem cells freeze-dried powder: total protein quantitative test box BCA method, freeze-dried powder one
Branch, which is dissolved in a solvent, to be sampled, and 5 times of dilutions, 6 measurement pipes, distilled water blank tube and standard pipe, microplate reader measure each pipe extinction
Value calculates total protein content, and the protein content of every freeze-dried powder should be in 1mg ± 0.05mg.
Human umbilical cord mesenchymal stem cells freeze-dried powder appearance and Detection of Stability: freeze-dried powder produced is white free from admixture powder
Last shape is faint yellow to colourless slightly thick liquid, clear after being dissolved with solvent.Special pH test paper detects dissolved freeze-drying
Powder, pH value is between 6.5-7.5.Freeze-dried powder saves at room temperature, and 1d/7d/14d appearance is unchanged；4 DEG C of preservations, 1d/ after dissolution
7d/14d appearance is unchanged.
The measurement of human umbilical cord mesenchymal stem cells secretion factor: ELISA kit is to two kinds of cell factor epidermal growth factors
The content of sub (EGF) and fibroblast growth factor (FGF) is measured, and the cell factor stoste of filtering and concentrating is taken to be surveyed
Fixed, it is 173ng/ml that experimental result, which can obtain EGF content, and FGF content is 246ng/ml, in acceptability limit.
The beneficial effects of the present invention are:
(1) present invention preferably source of the human umbilical cord mesenchymal stem cells as active cytokine, mescenchymal stem cell
Epidermal growth factor, vascular endothelial growth factor and stem cell factor isoreactivity factor component, these factor energy are rich in supernatant
Enough promote the growth and proliferation of internal stem cell, cell tissue in enteron aisle etc. is promoted to accelerate growth rate.It is pure by the present invention
The cell factor stoste purity is high of change, protein content is high, significant effect.
(2) present invention securely and reliably will not be as traditional using the culture medium culture umbilical cord stem cells containing serum substitute
Serum free culture system method may cause immunological rejection.
(3) dry powder is made in stem cell factor by the present invention, can be saved under cryogenic for a long time；Before use,
Deionized water and dry powder can be mixed and be used, solve the problems, such as that stem cell factor cannot save for a long time.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (9)
1. a kind of preparation method for repairing mammalian gut damage freeze-dried powder, which comprises the following steps:
(1) screening and culturing object and separate: the screening of umbilical cord is to choose pregnant woman age in 22-28 one full year of life, and pregnancy period≤40 week are healthy
Mature neonatal umbilical cord is placed in close to baby end about 20cm or so containing dual anti-umbilical cord acquisition liquid preservation, in ice chest as early as possible
Transport；
(2) it umbilical cord cleaning and processing: is placed in sterile cup ware with 75% alcohol Rapid Cleaning one time, then with containing dual anti-sterile life
Salt water repeated flushing is managed, until without obvious clot；Umbilical cord is longitudinally splitted, blood vessel is rejected；With containing dual anti-sterile saline
By tissue block rinsed clean, and divides segment to shred on umbilical cord with eye scissors and be placed in sterile cup ware, further shred umbilical cord
To fine tissue block；
(3) umbilical cord tissue culture: tissue block is inoculated in culture bottle bottom according to suitable density, adds suitable generation containing serum
For the DMEM/F12 culture solution of object, by body turnover bottle bottom think on be placed in 37 DEG C, adherent 4h in the incubator of 5% CO2, so
Overturning body bottom of bottle, which is statically placed in incubator downwards, afterwards cultivates, routine observation replacement or addition culture solution；8d or so is in microscope
Lower observation, it is seen that there is into fibrous cell around tissue block and climbs out of, removal tissue block replacement culture solution continues to cultivate 2d or so, this
When cell be P0, attached cell be transferred in new culture bottle cultivated with TrypLE digestion, cell is P1 at this time；
(4) mescenchymal stem cell secondary culture: the cell separated in step (3) is continued into secondary culture to P6, passage is when training
When cell growth reaches 80% fusion in feeding bottle, TrypLE vitellophag, 1: 4 secondary culture starts the culture for collecting P2 to P6
Supernatant；
(5) centrifugation of mescenchymal stem cell culture supernatant, purifying and filtration sterilization: the culture supernatant collected in (4) is carried out high
Speed centrifugation, again with supernatant after the centrifugation of sterile apyrogeneity container collection；By centrifuged supernatant concentration, then pass through filtering dress
It sets and degerming is carried out to concentrate and is dispensed into sterile apyrogeneity bottle；
(6) preparation of freeze-dried powder: KFG and freeze drying protectant is added in frozen dried concentrate in concentrate liquid；It is chilled dry
Mescenchymal stem cell factor freeze-dried powder is made after being dried.
2. a kind of preparation method for repairing mammalian gut damage freeze-dried powder according to claim 1, which is characterized in that
Using DMEM/F12 (HEPES, NO PHENOL RED, L-GLUTAMINE) the culture solution culture people's navel for containing 1% serum substitute
Band mescenchymal stem cell.
3. a kind of preparation method for repairing mammalian gut damage freeze-dried powder according to claim 1, which is characterized in that
The diameter dimension that umbilical cord scissors in step (1) is broken to fine tissue block is 2mm-4mm.
4. a kind of preparation method for repairing mammalian gut damage freeze-dried powder according to claim 1, which is characterized in that
The culture bottle of step (3) be T75cm2The surface sterile apyrogeneity TC vapor-permeable type culture bottle, the culture bottle for being passaged to P1 are
T175cm2The surface sterile apyrogeneity TC vapor-permeable type culture bottle.
5. a kind of preparation method for repairing mammalian gut damage freeze-dried powder according to claim 1, which is characterized in that
The culture supernatant collected in step (5) is to take centrifuged supernatant with 3000r/min centrifugation 30min, upper by what is repeatedly collected
Clear liquid is stored in -20 DEG C.
6. a kind of preparation method for repairing mammalian gut damage freeze-dried powder according to claim 1, which is characterized in that
The culture supernatant filtering and concentrating of step (5) is the culture supernatant that will collect in thaw at RT, passes through 0.45um membrane filtration
Participation cell is removed, molecular weight is being collected in the concentrate of the cell factor of 3~30KD by filter membrane ultrafiltration, is passing through 0.22um
Membrane filtration degerming, concentrate is collected.
7. a kind of preparation method for repairing mammalian gut damage freeze-dried powder according to claim 1, which is characterized in that
It is 0.01% that KFG ratio is added in step (6)；The freeze drying protectant ingredient include: trehalose, mannitol, dextran with
And human serum albumins.
8. a kind of preparation method for repairing mammalian gut damage freeze-dried powder according to claim 1, which is characterized in that
After freeze drying protectant is added in step (6), percent by volume of the trehalose in concentrate is 1~8%, and mannitol is in concentrate
In percent by volume be 1~8%, percent by volume of the dextran in concentrate be 1~5%, human serum albumins exists
Percent by volume in concentrate is 0.1~2%.
9. a kind of preparation method for repairing mammalian gut damage freeze-dried powder according to claim 1, which is characterized in that
Mescenchymal stem cell factor freeze-dried powder is made after chilled dry drying process in step (6), storage temperature is 2 DEG C -4
℃。
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110590932A (en) * | 2019-10-21 | 2019-12-20 | 南京先卓生物科技有限公司 | Preparation method of umbilical cord mesenchymal stem cell factor freeze-dried powder |
| CN112625114A (en) * | 2020-12-31 | 2021-04-09 | 瑞诺威德(杭州)医学科技有限公司 | Protein-containing protective agent and preservation method of mesenchymal stem cell factor |
| CN112707962A (en) * | 2020-12-31 | 2021-04-27 | 瑞诺威德(杭州)医学科技有限公司 | Carbohydrate-containing protective agent and preservation method of mesenchymal stem cell factor |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106367386A (en) * | 2016-10-14 | 2017-02-01 | 中卫华医（北京）生物科技有限公司 | Preparation method of human umbilical cord mesenchymal stem cell factor freeze-dried powder |
| CN108309822A (en) * | 2018-02-26 | 2018-07-24 | 福建省银丰干细胞工程有限公司 | A kind of preparation method of human umbilical cord mesenchymal stem cells paracrine factor freeze-dried powder |
| CN108823156A (en) * | 2018-07-04 | 2018-11-16 | 陕西神州生物技术有限公司 | For the clinical grade human umbilical cord mesenchymal stem cells composite factor of reparation and the preparation method of freeze-dried powder |
-
2018
- 2018-09-27 CN CN201811127256.3A patent/CN109010369A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106367386A (en) * | 2016-10-14 | 2017-02-01 | 中卫华医（北京）生物科技有限公司 | Preparation method of human umbilical cord mesenchymal stem cell factor freeze-dried powder |
| CN108309822A (en) * | 2018-02-26 | 2018-07-24 | 福建省银丰干细胞工程有限公司 | A kind of preparation method of human umbilical cord mesenchymal stem cells paracrine factor freeze-dried powder |
| CN108823156A (en) * | 2018-07-04 | 2018-11-16 | 陕西神州生物技术有限公司 | For the clinical grade human umbilical cord mesenchymal stem cells composite factor of reparation and the preparation method of freeze-dried powder |
Non-Patent Citations (2)
| Title |
|---|
| YUNBING WU ET AL.: "Exosomes derived from human umbilical cord mesenchymal stem cells alleviate inflammatory bowel disease in mice through ubiquitination", 《AMERICAN JOURNAL OF TRANSLATIONAL RESEARCH》 * |
| 张学娟 等: "无血清和有血清培养人脐带间充质干细胞的对比", 《中国组织工程研究》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110590932A (en) * | 2019-10-21 | 2019-12-20 | 南京先卓生物科技有限公司 | Preparation method of umbilical cord mesenchymal stem cell factor freeze-dried powder |
| CN112625114A (en) * | 2020-12-31 | 2021-04-09 | 瑞诺威德(杭州)医学科技有限公司 | Protein-containing protective agent and preservation method of mesenchymal stem cell factor |
| CN112707962A (en) * | 2020-12-31 | 2021-04-27 | 瑞诺威德(杭州)医学科技有限公司 | Carbohydrate-containing protective agent and preservation method of mesenchymal stem cell factor |
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Application publication date: 20181218 |