CN109464374A - Umbilical cord mesenchymal stem cells factor coniplexes are promoting the application in hair restoration - Google Patents
Umbilical cord mesenchymal stem cells factor coniplexes are promoting the application in hair restoration Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/14—Drugs for dermatological disorders for baldness or alopecia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q7/00—Preparations for affecting hair growth
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
Abstract
The present invention relates to cell factor purposes fields, are promoting the application in hair restoration in particular to umbilical cord mesenchymal stem cells factor coniplexes.It is a discovery of the invention that umbilical cord mesenchymal stem cells factor coniplexes can fundamentally activate hair follicle cell, promote hair restoration, significant effect, and it is more longlasting.
Description
Technical field
The present invention relates to cell factor purposes fields, exist in particular to umbilical cord mesenchymal stem cells factor coniplexes
Promote the application in hair restoration.
Background technique
Alopecia areata is a kind of non-cicatrical, inflammatory alopecia, and common clinical manifestation is that head the circle of sharpness of border occurs
Or oval plaque-like alopecia.There is whole alopecia or general alopecia in half conditions of patients recurrent exerbation, serious person, can delay the several years or tens of
Year.The characteristics of pelada and lipsotrichia is all hair follicle regresses, and hair follicle is malnutritive, and stand-down hair follicle increases, and hair follicle reduces, degenerates.At present
Essential therapeutic arsenals have using glucocorticoid, it is more effective to acute stage and the lesser pelada and lipsotrichia of area.
The treatment of pelada and lipsotrichia is extremely complex, needs to develop programs for each pathogenic factor, treatment method used at present
Mainly drug therapy.However hormone therapy is used, to body, other functions are affected, and only smaller to acute stage and area
It is effective.There are more and more people to attempt to promote hair regeneration, safety with cell factor now, and can fundamentally activate
Hair follicle cell, effect are more longlasting.
In view of this, the present invention is specifically proposed.
Summary of the invention
The purpose of the present invention is to provide a kind of umbilical cord mesenchymal stem cells factor coniplexes in promoting hair restoration
Using.
In order to realize above-mentioned purpose of the invention, the following technical scheme is adopted:
Umbilical cord mesenchymal stem cells factor coniplexes are promoting the application in hair restoration.
It is a discovery of the invention that umbilical cord mesenchymal stem cells factor coniplexes have the function of promoting in hair restoration, it is especially right
In alopecia areata.
Further, the umbilical cord mesenchymal stem cells factor coniplexes are prepared by the following method:
Umbilical cord mesenchymal stem cells carry out adhere-wall culture；
Liquid is changed, continues to cultivate, until cell is covered with to 85%-95%, culture solution is collected, obtains containing a large amount of umbilical cord mesenchymas
The culture solution of the factor of stem cell secretion；
The culture solution filtering removal cell fragment, obtains the liquid containing the umbilical cord mesenchymal stem cells factor coniplexes
Body.
The preparation method of umbilical cord mesenchymal stem cells factor coniplexes provided by the invention, after culture appropriate, cell
Factor content is abundant, and method is easy.
Further, the umbilical cord mesenchymal stem cells select P3-P7 for any one or more of.
P3-P7 is good for umbilical cord mesenchymal stem cells growth conditions, is easy culture, and activity is strong, what is be obtained by culture is thin
Intracellular cytokine content is more.
As in various embodiments, umbilical cord mesenchymal stem cells can select P3 generation, P4 generation, P5 generation, P6 generation, P7 generation etc.
Deng.
Further, the component for cultivating culture medium used is as follows:
On the basis of without phenol red α-MEM, add following components: FBS percentage by volume is 10% ± 2%, glutamine
Liquor capacity percentage is that 1% ± 0.05%, bFGF concentration is 20 ± 2ng/mL, and EGF concentration is 20 ± 2ng/mL；
Wherein, the concentration of the glutamine solution is 200mM.
The present invention selects the culture medium to cultivate umbilical cord mesenchymal stem cells, is conducive to umbilical cord mesenchymal stem cells and secretes
Trophic factors.
Further, the adhere-wall culture carries out in culture bottle, in terms of T75 culture bottle, is inoculated with 1.0 in each culture bottle
×106-2.0×106A cell, wherein the volume of the culture medium added is 8-12mL.
Further, condition of culture is equal are as follows: and 37 DEG C, 5%CO2, saturated humidity.
Culture carries out in the incubator.
Further, after the adhere-wall culture 8-15h, culture solution is sucked to remove not adherent cell and cell fragment,
Culture solution is rejoined, continues to cultivate.
Further, described to continue after cultivating 2-3 days, collect culture solution.Continue culture 2-3 days after, cell cover with to
90% or so, at this point, collecting culture solution, culture solution contains more cell factor.
Further, the filtering is carried out using not less than 20 μm of aperture sieve.
Preferably, it is carried out using 20-40 μm of aperture sieve.
After the screen to filtrate, remove cell debris or fragment and some cells not removed, it is dry to obtain umbilical cord mesenchyma
Cytokine complex.
Further, the liquid containing the umbilical cord mesenchymal stem cells factor coniplexes obtained further includes the step of packing
Suddenly, cryo-conservation is then carried out.Generally saved at -20 DEG C.
Mescenchymal stem cell (mesenchymal stem cells, MSC) be mesoderma origin, have height self more
The multipotential stem cell of new ability and multi-lineage potential.MSC is widely present in whole body Various Tissues, almost derives from body institute
Some histoorgans, such as marrow, periosteum, adipose tissue, dental pulp, synovial membrane, umbilical cord, placenta, amniotic fluid and fetal tissue.Difference is come
The mescenchymal stem cell in source has similar form, expresses identical surface markers, in terms of having similar biological characteristics, such as
Amplification can be cultivated in vitro, and can be divided under given conditions thin including nerve cell, osteoblast, cartilage cell, muscle
The cell of more organization systems including born of the same parents, fat cell.MSC is also making other than with multidirectional histocyte differentiation capability
Important regulative is played in blood, immunization inflammatory reaction, angiogenesis et al. body critical function.
Umbilical cord mesenchymal stem cells, which refer to, is present in one of neonatal umbilical cord tissue versatile stem cell.It is filled between umbilical cord
Matter is a kind of low immunogenicity cell, and has very strong immunoloregulation function.Umbilical cord mesenchymal stem cells possess the whole of MSC
Characteristic, and rich content are easily isolated culture.
Compared with the MSC in other sources, UC-MSC is more original, is dry between embryonic stem cell and adult stem cell
Cell, proliferation is lower than embryonic stem cell with differentiation capability, but is apparently higher than adult stem cell.
Umbilical cord mesenchymal stem cells have more differentiation potentials, and under specific inductive condition, UC-MSC can not only be divided into
The mesoblastemas such as bone, cartilage, fat, tendon, and can inwardly embryonic tissue cell (such as cardiac muscle cell, liver cell) and outside
Embryonic tissue cell (such as nerve cell) differentiation.UC-MSC does not have oncogenicity, can different inductive condition and it is suitable in vivo
It grows in microenvironment, safely directed differentiation is different tissue lines, has the ability for repairing various tissues and organ.
UC-MSC is exactly paracrine there are one important function other than it can be divided into the damage of histocyte repair tissue
Cell factor carries out tissue repair, such as secretory nerve Cell health factor, angiogenic factor, the hematopoiesis support factor, immune tune
Save the factor, anti-apoptosis factor and chemotactic factor (CF) etc..In the highly expressed factor of UC-MSC, G-CSF, GM-CSF, HGF, LIF, IL-
11 belong to Hemopoietic factor, there is promotion hemoposieis.HGF, IL-1 α, IL-6, IL-8 belong to immune-regulating factor.There are also EGF,
A variety of active factors that can adjust skin function such as FGF, VEGF, TGF-β, KGF.
Umbilical cord mesenchymal stem cells secretion promotes the hair restoration factor:
1) epidermal growth factor (EGF)
EGF is that the stimulation of epithelial cell division growth is former, and EGF activates a series of lifes in conjunction with the EGF receptor on cell membrane
Change reaction, change intracellular calcium concentration, starts gene related with cell division, so that akinete is entered division stage, have
Effect promotes and adjusts hair follicle cell growth, proliferation, promotes hair restoration.
2) fibroblast growth factor (FGF)
FGF is a kind of multi-functional nonspecific activity substance, and the various kinds of cell such as Human Umbilical Vein Endothelial Cells, fibroblast have promotion
Cell division effect.FGF can improve the microenvironment of cell growth, promote the growth and development of hair follicle cell.The rush new life function of FGF
It can, moreover it is possible to promote the reparation of impaired hair follicle.
3) keratinocyte growth factor (KGF)
KGF can promote epithelial cell proliferation and differentiation, moreover it is possible to keep cytoskeleton to stablize, organize the disconnected of actin
It splits, keeps cell results complete, avoid Apoptosis aging caused by active oxygen, impaired hair follicle cell can be repaired.
4) transforming growth factor β (TGF-β)
TGF-β is fibroblastic major chemokine, can stimulate fibroblast division growth.TGF-β at
The inducing chemotactic of fibrocyte is conducive to the progress of process of tissue reparation.TGF-β can also inducing macrophage secretion blood vessel hair
The raw factor, accelerates vascularization, and sufficient nutrition is provided for natural on-off cycles of hair growth.
5) vascular endothelial growth factor (VEGF)
VEGF is that presently found effect is most strong, the highest angiogenic factors of specificity.Scalp surface tissue is without blood
Pipe structure, it is main to provide nutriment by dermal microvascular and metabolic waste object is discharged.VEGF can effectively improve local vascular
Permeability provides sufficient nutriment and growth factor for fibroblastic proliferation and collage synthesis, promotes cell division,
Improve hair follicle microcirculation.
The preparation method of umbilical cord mesenchymal stem cells factor coniplexes provided by the invention, what is obtained contains umbilical cord mesenchyma
The liquid of stem cell factor compound is rich in above-mentioned substance.
Umbilical cord mesenchymal stem cells factor coniplexes provided by the invention fundamentally activate hair follicle cell, promote hair
Regeneration, significant effect, and it is more longlasting.
Liquid provided by the invention containing umbilical cord mesenchymal stem cells factor coniplexes, specifically used steps are as follows:
1) head is cleaned up with shampoo in advance, keeps scalp dry condition.
2) dedicated one layer of anaesthetic of skin is smeared, after smearing uniformly, one layer of covering preservative film leads to thickness to prevent anaesthetic flowing
Unevenly, 60-70min is waited.
3) anaesthetic is blown off with scraper plate, wipe residual anaesthetic with water gauze is dipped in.
4) operator puts on sterile gloves, opens sterile gauze, dips appropriate skin degerming bromogeramine, and wiping is de-
Hair, alopecia areata position twice, carry out disinfection.
5) remaining bromogeramine is wiped out with dry sterile gauze, until no moisture remains.
6) adjusting electronic micropin length is 1mm, carries out micropin needle thorn in scalp sites order, guarantees that every build skin all relates to
And there is micro bleeding category normal phenomenon.
7) the mescenchymal stem cell composite factor dissolved in advance is added dropwise simultaneously, with hand smear uniformly, make compound slowly by
It absorbs.
Compared with prior art, the invention has the benefit that
(1) umbilical cord mesenchymal stem cells factor coniplexes provided by the invention, cell factor type is abundant, and content is high, right
Hair restoration is promoted to have positive effect.
(2) umbilical cord mesenchymal stem cells factor coniplexes provided by the invention fundamentally activate hair follicle cell, promote head
Hair regeneration, significant effect, and it is more longlasting.
(3) preparation method of umbilical cord mesenchymal stem cells factor coniplexes provided by the invention, obtain containing between umbilical cord
The liquid of mesenchymal stem cells factor coniplexes is rich in EGF, FGF, KGF, TGF-β and VEGF.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described.
Fig. 1 is hair restoration variation comparison photo in experimental example 1 of the present invention；
Fig. 2 is another group of hair restoration variation comparison photo in experimental example 1 of the present invention.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
Embodiment 1
1, the preparation of umbilical cord mesenchymal stem cells culture medium
On the basis of without phenol red α-MEM, add following components: FBS percentage by volume is 10%, glutamine solution body
Product percentage is that 1%, bFGF concentration is 20ng/mL, and EGF concentration is 20ng/mL；
Wherein, the concentration of the glutamine solution is 200mM.
2, the preparation method containing umbilical cord mesenchymal stem cells factor coniplexes
1) it P3 generation, the good umbilical cord mesenchymal stem cells of growth conditions, is resuspended with complete medium.Each T75 bottles of inoculation
1.0×106A cell, complete medium dosage are 10mL.Jiggling is uniformly distributed cell, is steadily put into 37 DEG C, 5%CO2
It is overnight adherent in incubator.
2) culture solution is sucked within second day to remove not adherent cell and some cell fragments, rejoin 10mL and train completely
Base is supported, 37 DEG C, 5%CO are put into2It is cultivated in incubator.
3) visual cell's growth conditions, general 2-3 days cells are covered with to 90% or so, have contained a large amount of navels in culture solution at this time
The factor with mescenchymal stem cell secretion.The culture solution that each culture bottle is collected is focused in clean and sterile bottle.
4) culture solution of collection is filtered with 40 μm of cell screen clothes, removes cell fragment, detects component therein, as a result such as
Shown in table 1.
1 component situation of table
| EGF(ng/mL) | FGF(pg/mL) | KGF(ng/mL) | TGF-β(pg/mL) | VEGF(ng/mL) |
| 63.20±5.47 | 22.39±3.75 | 39.03±2.99 | 30.42±5.84 | 673.80±50.14 |
Obtained liquid is dispensed with sterile 5mL EP pipe, and -20 DEG C freeze, spare.
Embodiment 2
1, the preparation of umbilical cord mesenchymal stem cells culture medium
On the basis of without phenol red α-MEM, add following components: FBS percentage by volume is 10% ± 2%, glutamine
Liquor capacity percentage is that 1% ± 0.05%, bFGF concentration is 20 ± 2ng/mL, and EGF concentration is 20 ± 2ng/mL；
Wherein, the concentration of the glutamine solution is 200mM.
2, the preparation method containing umbilical cord mesenchymal stem cells factor coniplexes
1) it P4 generation, the good umbilical cord mesenchymal stem cells of growth conditions, is resuspended with complete medium.Each T75 bottles of inoculation
1.0×106A cell, complete medium dosage are 10mL.Jiggling is uniformly distributed cell, is steadily put into 37 DEG C, 5%CO2
It is overnight adherent in incubator.
2) culture solution is sucked within second day to remove not adherent cell and some cell fragments, rejoin 10mL and train completely
Base is supported, 37 DEG C, 5%CO are put into2It is cultivated in incubator.
3) visual cell's growth conditions, general 2-3 days cells are covered with to 90% or so, have contained a large amount of navels in culture solution at this time
The factor with mescenchymal stem cell secretion.The culture solution that each culture bottle is collected is focused in clean and sterile bottle.
4) culture solution of collection is filtered with 40 μm of cell screen clothes, removes cell fragment, detects component therein, as a result such as
Shown in table 2.
2 component situation of table
| EGF(ng/mL) | FGF(pg/mL) | KGF(ng/mL) | TGF-β(pg/mL) | VEGF(ng/mL) |
| 62.80±4.78 | 23.15±3.67 | 38.52±2.73 | 30.22±5.56 | 668.09±45.01 |
Obtained liquid is dispensed with sterile 5mL EP pipe, and -20 DEG C freeze, spare.
Embodiment 3
1, the preparation of umbilical cord mesenchymal stem cells culture medium
On the basis of without phenol red α-MEM, add following components: FBS percentage by volume is 10%, glutamine solution body
Product percentage is that 1%, bFGF concentration is 20ng/mL, and EGF concentration is 20ng/mL；
Wherein, the concentration of the glutamine solution is 200mM.
2, the preparation method containing umbilical cord mesenchymal stem cells factor coniplexes
1) it P7 generation, the good umbilical cord mesenchymal stem cells of growth conditions, is resuspended with complete medium.Each T75 bottles of inoculation
1.0×106A cell, complete medium dosage are 10mL.Jiggling is uniformly distributed cell, is steadily put into 37 DEG C, 5%
CO2, overnight adherent in saturated humidity incubator.
2) culture solution is sucked within second day to remove not adherent cell and some cell fragments, rejoin 10mL and train completely
Base is supported, 37 DEG C, 5%CO are put into2, cultivate in saturated humidity incubator.
3) visual cell's growth conditions, general 2-3 days cells are covered with to 90% or so, have contained a large amount of navels in culture solution at this time
The factor with mescenchymal stem cell secretion.The culture solution that each culture bottle is collected is focused in clean and sterile bottle.
4) culture solution of collection is filtered with 40 μm of cell screen clothes, removes cell fragment, detects component therein, as a result such as
Shown in table 3.
3 component situation of table
| EGF(ng/mL) | FGF(pg/mL) | KGF(ng/mL) | TGF-β(pg/mL) | VEGF(ng/mL) |
| 62.03±4.96 | 21.87±3.32 | 38.75±2.82 | 29.67±6.01 | 657.45±53.05 |
Obtained liquid is dispensed with sterile 5mL EP pipe, and -20 DEG C freeze, spare.
Comparative example 1
1) it is counted after digesting human umbilical cord mesenchymal stem cells, using mesenchymal stem cell serum-free culture medium by people's umbilical cord
Mescenchymal stem cell suspends, and adjustment cell concentration is 1.5 × 106A/mL.
2) by above-mentioned inoculation of suspension liquid into T75 culture bottle, culture overnight
3) it the liquid in culture bottle is absorbed, washs culture bottle with α-MEM, then add the α-MEM of 10mL, 6000U
RhPDGF-BB-the BB of recombinant human interferon alpha 2 and 600ng.
4) in 37 DEG C, 5%CO2, cultivate for 24 hours under the conditions of saturated humidity, then add the recombinant human epo of 60IU, continue
Culture 72 hours collects supernatant, obtains human umbilical cord mesenchymal stem cells culture supernatant.
5) culture solution of collection is filtered with 40 μm of cell screen clothes, removes cell fragment, detect component therein, cell because
The results are shown in Table 4 for the content detection of son.
4 component situation of table
| EGF(ng/mL) | FGF(pg/mL) | KGF(ng/mL) | TGF-β(pg/mL) | VEGF(ng/mL) |
| 35.34±3.25 | 12.15±4.73 | 20.08±2.56 | 15.12±5.78 | 200.45±37.54 |
Comparative example 2
It directly selects P4 to be cultivated for umbilical cord mesenchymal stem cells using the method for comparative example 1, obtains filling between people's umbilical cord
The filtering of matter stem cell culture supernatant, removes cell and its fragment, the results are shown in Table 5 for the content detection of cell factor.
5 component situation of table
| EGF(ng/mL) | FGF(pg/mL) | KGF(ng/mL) | TGF-β(pg/mL) | VEGF(ng/mL) |
| 38.044±2.97 | 16.42±3.78 | 24.12±2.46 | 16.01±4.38 | 270.45±32.15 |
Said components testing result finds that EGF, FGF, KGF in 1-3 of the embodiment of the present invention, TGF-β, VEGF component contain
No notable difference is measured, and EGF, FGF of comparative example 2, KGF, TGF-β, VEGF constituent content are slightly above comparative example 1.
Experimental example 1
Liquid containing umbilical cord mesenchymal stem cells factor coniplexes made from the above-mentioned preparation method of the present invention is to scalp
It is treated at position, the specific steps are as follows:
1) head is cleaned up with shampoo in advance, keeps scalp dry condition.
2) dedicated one layer of anaesthetic of skin is smeared, after smearing uniformly, one layer of covering preservative film leads to thickness to prevent anaesthetic flowing
Unevenly, 60-70min is waited.
3) anaesthetic is blown off with scraper plate, wipe residual anaesthetic with water gauze is dipped in.
4) operator puts on sterile gloves, opens sterile gauze, dips appropriate skin degerming bromogeramine, and wiping is de-
Hair, alopecia areata position twice, carry out disinfection.
5) remaining bromogeramine is wiped out with dry sterile gauze, until no moisture remains.
6) adjusting electronic micropin length is 1mm, carries out micropin needle thorn in scalp sites order, guarantees that every build skin all relates to
And there is micro bleeding category normal phenomenon.
7) the mescenchymal stem cell composite factor dissolved in advance is added dropwise simultaneously, with hand smear uniformly, make compound slowly by
It absorbs.
Male or female, the age 30-50 years old, patients with alopecia areata.The liquid provided using the embodiment of the present invention 1 treats 20 people altogether,
Each treatment interval time is 3 weeks, is treated 5 times altogether, and effect statistics is carried out.
Therapeutic effect discovery is counted, it is bald after the mescenchymal stem cell composite factor treatment provided through the embodiment of the present invention 1
Position grows denseer new hair, in addition, treating 2 times after the mescenchymal stem cell composite factor treatment that the embodiment of the present invention 1 provides
Afterwards i.e. it can be seen that hair is born.
The partial results for the mescenchymal stem cell composite factor therapeutic effect that the embodiment of the present invention 1 provides are as shown in Figs. 1-2.
Wherein, in Fig. 1 and Fig. 2, A is picture before treating, and B is picture after treatment.
Above content it is found that after umbilical cord mesenchymal stem cells factor coniplexes liquid use provided by the invention can faster and
Preferably activation scalp hair follicles promote hair restoration.
In addition, umbilical cord mesenchymal stem cells factor coniplexes liquid provided by the invention is applied to other kinds of alopecia,
Such as alopecia seborrheica, it may have good effect.
Although illustrate and describing the present invention with specific embodiment, it will be appreciated that without departing substantially from of the invention
Many other change and modification can be made in the case where spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (10)
1. umbilical cord mesenchymal stem cells factor coniplexes are promoting the application in hair restoration.
2. application according to claim 1, which is characterized in that the umbilical cord mesenchymal stem cells factor coniplexes by with
The preparation of lower section method:
Umbilical cord mesenchymal stem cells carry out adhere-wall culture；
Liquid is changed, continues to cultivate, until cell is covered with to 85%-95%, collects culture solution, is obtained dry thin containing a large amount of umbilical cord mesenchymas
The culture solution of the factor of intracrine；
The culture solution filtering removal cell fragment, obtains the liquid containing the umbilical cord mesenchymal stem cells factor coniplexes.
3. application according to claim 2, which is characterized in that the umbilical cord mesenchymal stem cells were selected in P3-P7 generation
It is any one or more.
4. application according to claim 2, which is characterized in that the component for cultivating culture medium used is as follows:
On the basis of without phenol red α-MEM, add following components: FBS percentage by volume is 10% ± 2%, glutamine solution
Percentage by volume is that 1% ± 0.05%, bFGF concentration is 20 ± 2ng/mL, and EGF concentration is 20 ± 2ng/mL；
Wherein, the concentration of the glutamine solution is 200mM.
5. application according to claim 4, which is characterized in that the adhere-wall culture carries out in culture bottle, is cultivated with T75
Bottle is counted, and 1.0 × 10 are inoculated in each culture bottle6-2.0×106A cell, wherein the volume of the culture medium added is 8-12mL.
6. application according to claim 5, which is characterized in that condition of culture is equal are as follows: 37 DEG C, 5%CO2, saturated humidity.
7. application according to claim 2, which is characterized in that after the adhere-wall culture 8-15h, suck culture solution to remove
Not adherent cell and cell fragment, rejoins culture solution, continues to cultivate.
8. application according to claim 7, which is characterized in that it is described to continue after cultivating 2-3 days, collect culture solution.
9. according to the described in any item applications of claim 2-8, which is characterized in that the filtering is using not less than 20 μm of aperture sieve
Net carries out, it is preferred to use 20-40 μm of aperture sieve carries out.
10. according to the described in any item applications of claim 2-8, which is characterized in that is obtained is dry containing the umbilical cord mesenchyma
The liquid of cytokine complex further includes the steps that packing, then carries out cryo-conservation.
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| CN110540958A (en) * | 2019-04-08 | 2019-12-06 | 启迪汉洱康（嘉兴）生物科技有限公司 | Preparation of umbilical cord mesenchymal stem cell secretory factor and application thereof in hair growth |
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| CN111840330A (en) * | 2020-08-03 | 2020-10-30 | 北京诺赛启研再生医学研究院有限公司 | Anti-hair loss repairing composition and application |
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| CN110540958A (en) * | 2019-04-08 | 2019-12-06 | 启迪汉洱康（嘉兴）生物科技有限公司 | Preparation of umbilical cord mesenchymal stem cell secretory factor and application thereof in hair growth |
| CN110540958B (en) * | 2019-04-08 | 2021-09-03 | 禾美生物科技(浙江)有限公司 | Preparation of umbilical cord mesenchymal stem cell secretory factor and application thereof in hair growth |
| CN110227057A (en) * | 2019-06-10 | 2019-09-13 | 北京恒峰铭成生物科技有限公司 | A kind of hair gel cream of hair regeneration and preparation method thereof |
| CN110642933A (en) * | 2019-10-08 | 2020-01-03 | 陕西中鸿科瑞再生医学研究院有限公司 | Preparation method of stem cell factor compound with anti-hair loss and hair growth effects |
| CN110642933B (en) * | 2019-10-08 | 2021-06-08 | 陕西中鸿科瑞再生医学研究院有限公司 | Preparation method of stem cell factor compound with anti-hair loss and hair growth effects |
| WO2021077812A1 (en) * | 2019-10-21 | 2021-04-29 | 江苏艾洛特医药研究院有限公司 | Umbilical cord mesenchymal stem cell factor freeze-dried powder, preparation method therefor and application thereof |
| CN110693909A (en) * | 2019-10-28 | 2020-01-17 | 陕西中鸿科瑞再生医学研究院有限公司 | Preparation of umbilical cord mesenchymal stem cell factor with hair growth effect |
| CN110878287A (en) * | 2019-12-05 | 2020-03-13 | 伯仕利生物科技发展（盐城）有限公司 | Preparation method and application of stem cell supernatant over-expressing TAT-KGF |
| CN111557896A (en) * | 2020-06-23 | 2020-08-21 | 华悦汇(上海)健康科技有限公司 | Preparation method of cell factor for hair follicle activation function |
| CN111840330A (en) * | 2020-08-03 | 2020-10-30 | 北京诺赛启研再生医学研究院有限公司 | Anti-hair loss repairing composition and application |
| CN114681486A (en) * | 2020-12-30 | 2022-07-01 | 杭州易文赛生物技术有限公司 | Stem cell factor composition and application thereof in hair regeneration |
| CN113750116A (en) * | 2021-10-22 | 2021-12-07 | 广东唯泰生物科技有限公司 | A preparation for promoting hair growth, and its preparation method |
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