CN107714727A - A kind of enriching fat stem cell culture supernatant preparation, its preparation method and application - Google Patents
A kind of enriching fat stem cell culture supernatant preparation, its preparation method and application Download PDFInfo
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- CN107714727A CN107714727A CN201711003978.3A CN201711003978A CN107714727A CN 107714727 A CN107714727 A CN 107714727A CN 201711003978 A CN201711003978 A CN 201711003978A CN 107714727 A CN107714727 A CN 107714727A
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- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
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Abstract
The invention belongs to medical sanitary technology field, specifically provide a kind of enriching fat stem cell culture supernatant preparation for treating skin scald, mainly by adipose tissue-derived mescenchymal stem cell culture cell culture fluid supernatant concentration and obtain, also added with cell factor hrEGF, VEGF and bFGF.The present invention further correspondingly provides preparation method and its application of the enriching fat stem cell culture supernatant preparation.The enriching fat stem cell culture supernatant preparation of the present invention has obvious promotion repair for scald wound, is embodied in：Can faster it be repaired compared with positive controls and other embodiments group, epidermis and corium with recovering function after use.Also, living cells composition is free of in the enriching fat stem cell culture supernatant preparation of the present invention, in the absence of ethics or immune safety problem, and without matching while using, using convenient.
Description
Technical field
The invention belongs to medical sanitary technology field, and in particular to a kind of enriching fat stem cell culture supernatant preparation, its
Preparation method and application.
Background technology
The tissue damage as caused by the chemical substance such as the physical factors such as flame, hydrothermal solution, electricity and strong acid, highly basic, which is referred to as burning, scalds
Wound, it is daily life and the common disease in work.Deep tissue can be caused downright bad when scalding serious, if dealt with improperly, sternly
It can fester, can not all heal for a long time again.
Wound tissue healing process mainly includes inflammatory reaction phase, cellular proliferative stage and structure remodeling phase.After tissue damage,
Inflammatory engenders the propagation of fibroblast and capillary endothelial cell after oozing out, newborn with the formation of granulation tissue
Epithelium promotes to surface of a wound center, gradually fills up wound flap coverage until wound healing.
Mainly include currently for the processing means after scald：Physical temperature-lowering, surface of a wound sterilization are with sterilizing and using antibiotic
Prevention infection.Obviously, these processing means only belong to the precautionary measures of acute stage, wound repair after scald can not be produced straight
Connect effect.What is more, and the condition of the injury can be aggravated by dealing with improperly.For example, adverse reaction caused by rinsing cooling possibility with flowing water includes：
Cause to be complicated by infection by flowing water bacterium, make wound dehiscence or shrunken skin because of fluviation, scar is left after healing, because body temperature drops
It is low excessively to cause shock etc..
Also paid pilot production carries out repairing and treating to the surface of a wound to existing medical procedure with stem cell, but existing this therapeutic modality
Need to keep stem cell normal growth, therefore usually need matching while using to put, it is difficult to guarantee the quality, using being very limited.
In summary, the existing reparation means for scald wound are unreasonable, and adverse reaction is more, cumbersome, using by
Limit.Therefore it is a kind of to have good result to skin scald wound repair and treatment preparation easy to use is urgently developed and ground
Study carefully.
The content of the invention
In order to solve above mentioned problem existing for prior art, the present invention provides a kind of enriching fat stem cell culture supernatant system
Agent, the present invention seeks to strongly facilitate mitosis by the bioactivity of substantial amounts of cell factor and participate in stem cell induction point
Change, then participate in histiocytic growth, regeneration and rebuild, quickly complete the reparation for the surface of a wound.It is also corresponding in the present invention
Offer the enriching fat stem cell culture supernatant preparation preparation method and application.
According to the enriching fat stem cell culture supernatant preparation of the embodiment of the present invention, the enriching fat is dry thin
Born of the same parents' culture supernatant preparation mainly by adipose tissue-derived mescenchymal stem cell culture cell culture fluid supernatant concentration and
.
According to the enriching fat stem cell culture supernatant preparation of the embodiment of the present invention, preferable scheme is institute
State enriching fat stem cell culture supernatant preparation and be also added with hrEGF, VEGF and bFGF.It is further preferred that hrEGF, VEGF
Concentration after being added with bFGF is each followed successively by 0.08~0.15 μ g/mL, 0.05~0.11 μ g/mL and 0.01~0.10 μ respectively
g/mL。
According to the enriching fat stem cell culture supernatant preparation of the embodiment of the present invention, preferable scheme is institute
The multiple for stating concentration is 3~6 times, and the method for the concentration is dialysis.
According to the embodiment of the present invention, it is a further object of the present invention to provide enriching fat stem cell culture
The preparation method of Shangqing ', comprises the following steps：S1 takes adipocyte, is cleaned using physiological saline, then add etc.
The collagenase digesting of volume, centrifuged after digestion, remove supernatant；S2 inserts fat stem cell serum free medium, is transferred to after resuspension
Tissue Culture Flask is cultivated；S3 collects the supernatant before passage, and cell culture fluid will be made after supernatant concentration, adds
HrEGF, VEGF and bFGF are produced.
According to the present invention embodiment enriching fat stem cell culture supernatant preparation preparation method, preferably
Scheme is, in S1, the clostridiopetidase A is type i collagen enzyme, and time of the digestion is 30~60min, the type i collagen enzyme it is dense
Spend for 0.1%~0.5%.
According to the present invention embodiment enriching fat stem cell culture supernatant preparation preparation method, preferably
Scheme is, in S1, the centrifugation is specially：By postdigestive adipocyte with a diameter of 70 μm of membrane filtration, then
5~10min is centrifuged under conditions of 2000rpm/min.
According to the present invention embodiment enriching fat stem cell culture supernatant preparation preparation method, preferably
Scheme is, in S2, is added with hrEGF, VEGF and bFGF in the fat stem cell serum free medium, hrEGF, VEGF and
The concentration in culture medium after bFGF additions is respectively 0.08~0.15 μ g/mL, 0.05~0.11 μ g/mL and 0.01~0.10 μ
g/mL。
According to the present invention embodiment enriching fat stem cell culture supernatant preparation preparation method, preferably
Scheme is, degerming through 0.22 μm of membrane filtration of diameter after concentration in S3, gained enriching fat after addition hrEGF, VEGF and bFGF
HrEGF, VEGF and bFGF concentration are respectively 0.08~0.15 μ g/mL, 0.05~0.11 μ g/ in stem cell culture supernatant preparation
ML and 0.01~0.10 μ g/mL.
According to the embodiment of the present invention, another object of the present invention is to provide the enriching fat stem cell culture
Application of the Shangqing ' in the medicine for preparing treatment scald.Preferably, it is described scald include but is not limited to flame, hydrothermal solution,
Tissue damage caused by the chemical substance such as the physical factors such as electricity and strong acid and highly basic.The medicine includes but is not limited to liquid
Agent, pulvis, mist agent, pill and stick the formulations such as ointment.
In the present invention, fat stem cell can secrete cytokine profiles through in vitro culture, such as VEGF, HGF, bFGF, IGF-
I, SDF-1 α etc..The main biological activity of these cell factors is to strongly facilitate mitosis and participate in stem cell induction to divide
Change, it is thus possible to participate in histiocytic growth, regeneration and rebuild.
The specific of the present invention has the beneficial effect that：
First, the core content that the present invention is scalded from the concentration culture supernatant preparation of fat stem cell as treatment, energy
It is enough efficiently and effectively to repair scald wound, and repair process side effect is low, quick, has filled up after directly facilitating scald on the market
The vacancy that the externally applied drug of wound repair lacks.
Second, the enriching fat stem cell culture supernatant preparation of the present invention promotes scald wound repairing effect obvious, with
RhEGF positive Control examples, cycles of concentration are compared with two different comparative examples of cytokine concentrations, and after 7d, surface of a wound epidermis is complete
It is whole and thick, and had secretory cell appearance, and secretion capacity is strong；Subcutaneous collagen aligned orderly；Hair follicle, revascularization rule
It is and more.
Third, the enriching fat stem cell culture supernatant preparation of the present invention derives from people, no kind problem；And in finished product
Without living cells composition, in the absence of ethics or immune safety problem, and without matching while using, using convenient.
Fourth, the culture medium that the enriching fat stem cell culture supernatant preparation of the present invention is used when cultivating fat stem cell
Middle addition cell factor, it is relatively beneficial to the efficient reparation of the surface of a wound after scald.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
There is the required accompanying drawing used in technology description to be briefly described, it should be apparent that, drawings in the following description are only this
Some embodiments of invention, for those of ordinary skill in the art, on the premise of not paying creative work, can be with
Other accompanying drawings are obtained according to these accompanying drawings.
Fig. 1 uses 40 times of H＆E stained slices of skin after the treatment of the embodiment of the present invention 1 7d for skin after scald.
Fig. 2 uses 40 times of H＆E stained slices of skin after hrEGF positive Control examples treatment 7d for skin after scald.
Fig. 3 uses 40 times of H＆E stained slices of skin after the treatment of the embodiment of the present invention 2 7d for skin after scald.
Fig. 4 uses 40 times of H＆E stained slices of skin after the treatment of example one 7d in the embodiment of the present invention 3 for skin after scald.
Fig. 5 uses 40 times of H＆E stained slices of skin after the treatment of example two 7d in the embodiment of the present invention 3 for skin after scald.
Fig. 6 uses 40 times of H＆E stained slices of skin after the treatment of the embodiment of the present invention 4 7d for skin after scald.
Embodiment
To make the object, technical solutions and advantages of the present invention clearer, technical scheme will be carried out below
Detailed description.Obviously, described embodiment is only part of the embodiment of the present invention, rather than whole embodiments.Base
Embodiment in the present invention, those of ordinary skill in the art are resulting on the premise of creative work is not made to be owned
Other embodiment, belong to the scope that the present invention is protected.
Embodiment 1：
The present embodiment provides a kind of enriching fat stem cell culture supernatant preparation, and it is mainly by between adipose tissue-derived
The supernatant concentration of the cell culture fluid of mesenchymal stem cells culture and obtain.The enriching fat stem cell culture supernatant system of the present embodiment
The specific preparation method of agent is：Adipose tissue 40ml (adipose tissue-derived is preferably derived from people in animal) is taken, adding concentration is
0.1%~0.5% type i collagen 30~60min of enzymic digestion, after brine, under conditions of 2000rpm/min from
5~10min of the heart, takes sedimentation cell, and resuspension is cleaned with physiological saline, and with a diameter of 70 μm of membrane filtration, it is dry thin to obtain fat
With hrEGF, VEGF and bFGF of addition GMP standards, (concentration respectively is 0.08~0.15 μ g/mL, 0.05~0.11 μ g/ to born of the same parents
ML and 0.01~0.10 μ g/mL) fat stem cell serum free medium cultivated；Fat stem cell in the present embodiment without
Blood serum medium is in CO2Content is 5% moist aseptic gas environment, and temperature is to be cultivated in 37 DEG C of incubator.Cell is trained
The fat stem cell for being passaged to the third generation is supported through phenotypic evaluation, CD49d, CD105 are expressed as the positive, and CD106 is feminine gender, into fat
Osteogenic induction experiment confirms that it has the speciality of multi-lineage potential cell.Cell culture collects cell life to the three~five generation
For length to supernatant during 90% Fusion Strain, 3~6 times of dialysis concentration is simultaneously degerming through 0.22 μm of membrane filtration of diameter, finally adds
(final concentration respectively is 0.08~0.15 μ g/mL, 0.05~0.11 μ g/ to hrEGF, VEGF and bFGF of GMP standards after addition
ML and 0.01~0.10 μ g/mL) finished product is made.
Embodiment 2：
The present embodiment provides a kind of enriching fat stem cell culture supernatant preparation, and the difference with embodiment 1 is, this implementation
Example enriching fat stem cell culture supernatant preparation final step in eliminate using addition cell factor hrEGF, VEGF with
The step of bFGF.
The present embodiment concretely comprises the following steps：Adipose tissue 40ml is taken, adds the type i collagen enzyme that concentration is 0.1%~0.5%
30~60min is digested, after brine, 5~10min is centrifuged under conditions of 2000rpm/min, takes sedimentation cell,
Clean resuspension with physiological saline, with a diameter of 70 μm of membrane filtration, obtain fat stem cell addition GMP standards hrEGF,
VEGF and bFGF (concentration is respectively 0.08~0.15 μ g/mL, 0.05~0.11 μ g/mL and 0.01~0.10 μ g/mL) fat
Stem cell serum-free culture medium is cultivated；Fat stem cell serum free medium in the present embodiment is in CO2Content is 5%
Moist aseptic gas environment, temperature are to be cultivated in 37 DEG C of incubator.Cell culture passages to the third generation fat stem cell
Through phenotypic evaluation, CD49d, CD105 are expressed as the positive, and CD106 is feminine gender, and it is multidirectional to confirm that it has into the experiment of fat osteogenic induction
The speciality of differentiation potential cell.Cell culture collects supernatant during cell growth to 90% Fusion Strain to the three~five generation,
Simultaneously finished product is made through 0.22 μm of membrane filtration of diameter is degerming in 3~6 times of dialysis concentration.
Embodiment 3：
The present embodiment provides a kind of enriching fat stem cell culture supernatant preparation, and the difference with embodiment 1 is, finished product system
HrEGF, the VEGF for the GMP standards finally added in standby are different from bFGF and embodiment 1.The present embodiment 3 is divided to for two examples, point
It is not as follows：
Example one：This example one concretely comprises the following steps：Adipose tissue 40ml is taken, adds the type i collagen enzyme that concentration is 0.3%
30~60min is digested, after brine, 5~10min is centrifuged under conditions of 2000rpm/min, takes sedimentation cell,
Clean resuspension with physiological saline, with a diameter of 70 μm of membrane filtration, obtain fat stem cell addition GMP standards hrEGF,
(concentration is respectively that concentration is respectively 0.08~0.15 μ g/mL, 0.05~0.11 μ g/mL and 0.01~0.10 μ g/ to VEGF with bFGF
ML fat stem cell serum free medium)；Fat stem cell serum free medium in the present embodiment is in CO2Content is
5% moist aseptic gas environment, temperature are to be cultivated in 37 DEG C of incubator.The fat of cell culture passages to the third generation is done
Cell is through phenotypic evaluation, and CD49d, CD105 are expressed as the positive, and CD106 is feminine gender, confirms that it has into the experiment of fat osteogenic induction
The speciality of multi-lineage potential cell.Cell culture is upper when collecting cell growth to 90% Fusion Strain to the three~five generation
Clear liquid, dialysis concentration 3~5 times simultaneously it is degerming through 0.22 μm of membrane filtration of diameter, finally add GMP standards hrEGF, VEGF with
Finished product is made in bFGF (final content concn respectively is 0.25 μ g/mL, 0.15 μ g/mL and 0.20 μ g/mL).
Example two：This example two concretely comprises the following steps：Adipose tissue 40ml is taken, adds the type i collagen enzyme that concentration is 0.3%
30~60min is digested, after brine, 5~10min is centrifuged under conditions of 2000rpm/min, takes sedimentation cell,
Clean resuspension with physiological saline, with a diameter of 70 μm of membrane filtration, obtain fat stem cell addition GMP standards hrEGF,
(concentration is respectively that concentration is respectively 0.08~0.15 μ g/mL, 0.05~0.11 μ g/mL and 0.01~0.10 μ g/ to VEGF with bFGF
ML fat stem cell serum free medium) is cultivated；Fat stem cell serum free medium in the present embodiment is in CO2
Content is 5% moist aseptic gas environment, and temperature is to be cultivated in 37 DEG C of incubator.Cell culture passages are to the third generation
Fat stem cell is through phenotypic evaluation, and CD49d, CD105 are expressed as the positive, and CD106 is feminine gender, and being tested into fat osteogenic induction confirms
It has the speciality of multi-lineage potential cell.Cell culture collects cell growth to 90% Fusion Strain to the three~five generation
When supernatant, dialysis concentration 3~5 times simultaneously it is degerming through 0.22 μm of membrane filtration of diameter, finally add GMP standards hrEGF,
Finished product is made in VEGF and bFGF (final content concn respectively is 0.025 μ g/mL, 0.01 μ g/mL and 0.005 μ g/mL).
Embodiment 4：
The present embodiment provides a kind of enriching fat stem cell culture supernatant preparation, with differing only in for embodiment 1, removes
The step of dialysis concentration.
Embodiment 5：
The present embodiment provides a kind of enriching fat stem cell culture supernatant preparation, including four parallel tests, with embodiment 1
Differ only in, in four parallel tests finally addition GMP standards hrEGF final concentration be followed successively by respectively 0.08 μ g/mL,
0.10 μ g/mL, 0.12 μ g/mL and 0.15 μ g/mL.
Embodiment 6：
The present embodiment provides a kind of enriching fat stem cell culture supernatant preparation, including four parallel tests, with embodiment 1
Differ only in, in four parallel tests finally addition GMP standards bFGF final concentration be followed successively by respectively 0.01 μ g/mL,
0.04 μ g/mL, 0.07 μ g/mL and 0.10 μ g/mL.
Embodiment 7：
The present embodiment provides a kind of enriching fat stem cell culture supernatant preparation, including four parallel tests, with embodiment 1
Differ only in, in four parallel tests finally addition GMP standards VEGF final concentration be followed successively by respectively 0.05 μ g/mL,
0.06 μ g/mL, 0.08 μ g/mL and 0.11 μ g/mL.
Rat is tested using the finished product obtained by above-described embodiment 1~7, test method is as follows：
First, the deep two degree of scalding models of rat skin are established
By the way of 1. rat is after the chloral hydrate anesthesia, is scalded 10 seconds using 98 DEG C water steam, reach scald
It is deep II degree.
2. treatment method：Every group in triplicate.Twice daily, take quantitative medicine to be applied to scald surface, gently loop
To fully absorbing, 7d is used continuously, therapeutic process observes skin infection and repairs situation, and rat diet action etc. is other anti-
Should.
3. therapeutic effect detects
Treatment time puts to death rat after terminating, and takes skin holostrome and subsidiary part hypodermis to send pathological section, per a
Sample carries out H＆E dyeing；Testing result is as follows：
The epidermis of 3.1 embodiment 1 is complete and thick, has had secretory cell generation, and secretion capacity is strong；Collagen aligned orderly；
There is a hair follicle, blood vessel rule growth and more；Subcutaneous collagen generates and marshalling, and effect is compared with hrEGF groups and embodiment 2 and 3
It is all good；HrEGF control groups：Epidermis complete collyriculum, subcutaneous collagen arrangement disorder；Negative control group：Epidermis defect, collagen are disorderly
Disorderly；Embodiment 2：Epithelial cell not enough continues, and quantity is few, and collagen layer relatively lacks, and inflammatory infiltration concentrates on part, new vessels
Quantity is more, a little hair follicle, prompts to form refractory conjunction scar；Example one and two in embodiment 3：Epithelium is completely continuous, but secretes
Cell is less, and collagen marshalling, inflammatory cell infiltration, blood vessel is more, no hair follicles outgrowth, prompts to form granulation tissue；Conclusion：
Embodiment 1 has the function of good promotion scald wound reparation, and positive effect is better than remaining control group.Treat whole rabbit drink
Food action is normal.
3.2 sort out reparation situation table according to the H＆E results dyed, and reparation situation is as shown in table 1, in table 1+represent
Reparation degree ,+more, repairing effect is better, and-representative does not have reparation.
Table 1
| Epidermis | Collagen | Inflammatory infiltration | Blood vessel | Hair follicle | |
| Embodiment 1 | ++++ | ++++ | - | ++ | ++ |
| hrEGF | - | + | + | - | ++ |
| Embodiment 2 | + | + | +++ | +++ | + |
| The example one of embodiment 3 | +++ | +++ | ++ | +++ | - |
| The example two of embodiment 3 | ++ | ++ | ++ | +++ | - |
Note：The degree of tissue repair after how much representatives treatment of "+".
In summary：By the data in accompanying drawing 1~5 and table 1, the result of embodiment 2 and 3 can be proved only
Have and can be only achieved best repairing effect, two examples and the phase of embodiment 2 of embodiment 3 in the concentration range value of the present invention
It is mutually more visible, tissue repair effect corresponding to rhEGF, VEGF and bFGF embodiment 1 is not added most with the final finished stage
Difference, it is relatively preferable with tissue repair effect corresponding to the example two in the embodiment 3 of higher addition concentration, but all without this hair
Good, the of the invention cell culture fluid of tissue repair effect in bright preset range under cell factor addition concentration corresponding to embodiment
The effect mutually promoted between rhEGF, VEGF and bFGF be present, and each content has certain requirement for its.This hair
Bright effect is very notable.
Contrast accompanying drawing 1 and understand that the therapeutic effect in accompanying drawing 1 is obviously improved compared to accompanying drawing 6 with accompanying drawing 6.Show reality
Apply after concentration step processing has been carried out in example 1, have to the therapeutic effect of scald and be obviously improved.
Four parallel tests in embodiment 5 have carried out influence of the hrEGF ultimate densities to therapeutic effect, four parallel examinations
Test and carry out effect comparison according to foregoing rat test mode, analysis result is listed in the table below 2.
Table 2
Note：How much representatives of "+" represent the degree of tissue repair after treatment.
Analytical table 2 understands that hrEGF can be played pre- when predetermined concentration scope (0.08 μ g/mL~0.15 μ g/mL) is interior
Fixed scald repairing and treating effect, and when concentration is higher in the range of for hrEGF concentration, treatment repairing effect is better.
Four parallel tests in embodiment 6 have carried out influence of the bFGF ultimate densities to therapeutic effect, four parallel examinations
Test and carry out effect comparison according to foregoing rat test mode, analysis result is listed in the table below 3.
Table 3
Note：How much representatives of "+" represent the degree of tissue repair after treatment.
Analytical table 3 understands that bFGF can be played predetermined when predetermined concentration scope (0.06 μ g/mL~0.15 μ g/mL) is interior
Scald repairing and treating effect, and when bFGF concentration is in 0.13 μ g/mL, treatment repairing effect is best, particularly with epidermis
Repair, improve a lot.
Four parallel tests in embodiment 7 have carried out influence of the VEGF ultimate densities to therapeutic effect, four parallel examinations
Test and carry out effect comparison according to foregoing rat test mode, analysis result is listed in the table below 4.
Table 4
Note：How much representatives of "+" represent the degree of tissue repair after treatment.
Analytical table 4 understands that VEGF can be played predetermined when predetermined concentration scope (0.05 μ g/mL~0.11 μ g/mL) is interior
Scald repairing and treating effect, and when VEGF concentration is in 0.08 μ g/mL, treatment repairing effect is best, particularly with blood vessel
Palingenesis, improve a lot.
The foregoing is only a specific embodiment of the invention, but protection scope of the present invention is not limited thereto, any
Those familiar with the art the invention discloses technical scope in, change or replacement can be readily occurred in, should all be contained
Cover within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.
Claims (10)
- A kind of 1. enriching fat stem cell culture supernatant preparation, it is characterised in that：The enriching fat stem cell culture supernatant system Agent mainly by adipose tissue-derived mescenchymal stem cell culture cell culture fluid supernatant concentration and obtain.
- 2. enriching fat stem cell culture supernatant preparation according to claim 1, it is characterised in that：Also added with hrEGF, VEGF and bFGF.
- 3. enriching fat stem cell culture supernatant preparation according to claim 2, it is characterised in that：The multiple of the concentration For 3~6 times, the method for the concentration is dialysis.
- A kind of 4. preparation method of enriching fat stem cell culture supernatant preparation described in Claims 2 or 3, it is characterised in that bag Include following steps：S1 takes adipocyte, is cleaned using physiological saline, then adds isometric collagenase digesting, is centrifuged after digestion, Remove supernatant；S2 inserts fat stem cell serum free medium, and being transferred to Tissue Culture Flask after resuspension is cultivated；S3 collects the supernatant before passage, and cell culture fluid will be made after supernatant concentration, and addition hrEGF, VEGF and bFGF are .
- 5. preparation method according to claim 4, it is characterised in that：In S1, the clostridiopetidase A is type i collagen enzyme, described The time of digestion is 30~60min, and the concentration of the type i collagen enzyme is 0.1%~0.5%.
- 6. preparation method according to claim 5, it is characterised in that：In S1, the centrifugation is specially：By postdigestive fat A diameter of 70 μm of membrane filtration of fat cell, then 5~10min is centrifuged under conditions of 2000rpm/min.
- 7. preparation method according to claim 5, it is characterised in that：In S2, the fat stem cell serum free medium In be added with hrEGF, VEGF and bFGF, the concentration in culture medium after hrEGF, VEGF and bFGF addition is respectively 0.08~ 0.15 μ g/mL, 0.05~0.11 μ g/mL and 0.01~0.10 μ g/mL.
- 8. preparation method according to claim 4, it is characterised in that：In S3, through 0.22 μm of membrane filtration of diameter after concentration It is degerming, hrEGF, VEGF and bFGF in enriching fat stem cell culture supernatant preparation obtained by after addition hrEGF, VEGF and bFGF Concentration is respectively 0.08~0.15 μ g/mL, 0.05~0.11 μ g/mL and 0.01~0.10 μ g/mL.
- 9. a kind of any enriching fat stem cell culture supernatant preparation of claims 1 to 3 is preparing the medicine for the treatment of scald In application.
- 10. application according to claim 9, it is characterised in that the scald includes flame, hydrothermal solution, electricity, strong acid and highly basic Caused tissue damage, the medicine include liquor, pulvis, mist agent and stick ointment.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108478611A (en) * | 2018-04-25 | 2018-09-04 | 常州市阿曼特医药科技有限公司 | A kind of reparation medicament for treating scald |
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| CN110464878A (en) * | 2019-09-10 | 2019-11-19 | 深圳至博生物科技有限公司 | A kind of application of VEGF |
| CN112111451A (en) * | 2020-11-23 | 2020-12-22 | 北京欣颂生物科技有限公司 | Method for increasing yield of stem cell cytokines |
| CN114948998A (en) * | 2022-06-01 | 2022-08-30 | 复旦大学附属中山医院 | Polysaccharide biomedical colloidal fluid rich in human adipose-derived mesenchymal stem cell factor compound and preparation method and application thereof |
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