CN110151678A - It is a kind of for improving the composition and cosmetics comprising source of human stem cell excretion body of wrinkle - Google Patents
It is a kind of for improving the composition and cosmetics comprising source of human stem cell excretion body of wrinkle Download PDFInfo
- Publication number
- CN110151678A CN110151678A CN201910553006.4A CN201910553006A CN110151678A CN 110151678 A CN110151678 A CN 110151678A CN 201910553006 A CN201910553006 A CN 201910553006A CN 110151678 A CN110151678 A CN 110151678A
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- China
- Prior art keywords
- stem cell
- excretion body
- wrinkle
- cell excretion
- source
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Classifications
-
- 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
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- 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
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- 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
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Zoology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a kind of for improving the composition comprising source of human stem cell excretion body of wrinkle, includes adipose-derived mescenchymal stem cell excretion body and plant extracts.Adipose-derived mescenchymal stem cell excretion body is added in composition of the present invention, adipose-derived mescenchymal stem cell excretion body has the activity for inhibiting elastoser and Collagenase, it is effective against aging, improve the elasticity of skin, improve wrinkle of skin, and adipose-derived mescenchymal stem cell excretion body can also promote the absorption of plant extracts to a certain extent, keep the utilization rate of plant extracts in skin higher.The invention also discloses the cosmetics comprising the composition comprising source of human stem cell excretion body for being used to improve wrinkle.
Description
Technical field
The present invention relates to a kind of stem cell excretion bodies and application thereof, and in particular to a kind of for improving the thin comprising doing of wrinkle
The composition and cosmetics of born of the same parents source excretion body.
Background technique
Excretion body (Exosome) is by intracellular vesicles structure release to a kind of extracellular film property vesica, diameter about 30-
150nm, cell can all secrete excretion body under normal physiological status, due to merging between plasma membrane and multivesicular body as a result, outer
It secretes body sample vesica and external space of cell is secreted by exocytosis.Excretion body sample vesica be divided by double-layer of lipoid it is inside and outside,
Since it contains membrane lipid, memebrane protein, inhereditary material and cytoplasmic components, reflect the property and state of cell indirectly.Together
When, excretion body sample vesica by and other cells and tissue combination, and by film component, mRNA, miRNA, protein (growth swash
Element, cell factor etc.) etc. be transferred to recipient cell, and as the extracellular transport protein communicated between mediate cell-cell.
Skin aging is the damage collective effect such as the intrinsic physiological mechanism of human senility and extraneous exposing to the weather, environmental pollution
Result.Skin aging is usually embodied in the generation of skin elasticity reduction and wrinkle of skin, it was reported that collagen and elastic egg
White reduction will lead to skin elasticity reduction and wrinkle of skin generates, and collagen and elastin laminin are by Collagenase and elasticity
Protease regulator, Collagenase and elastoser can cause collagen and elastin degradation, so as to cause collagen
Albumen and elastin laminin are reduced, and wrinkle is caused to generate, and skin elasticity lowers.There has been no to adipose-derived mescenchymal stem cell at present
Correlative study of the excretion body in terms of skin wrinkle resisting anti-aging.
Summary of the invention
It is provided a kind of for improving the packet of wrinkle it is an object of the invention to overcome the shortcomings of the prior art place
The composition and cosmetics of the body of excretion containing source of human stem cell.
To achieve the above object, the technical scheme adopted by the invention is as follows: it is a kind of for improve wrinkle comprising stem cell come
The composition of source excretion body includes adipose-derived mescenchymal stem cell excretion body and plant extracts.Adipose-derived mesenchyma is dry
Cell excretion body has the activity for inhibiting elastoser and Collagenase, is effective against aging, improves the elasticity of skin,
Improve wrinkle of skin, and adipose-derived mescenchymal stem cell excretion body can also promote plant extracts to a certain extent
Absorption, keep the utilization rate of plant extracts in skin higher.
Preferably, the plant extracts includes gadol extract, Gotu Kola P.E and lupin extract.
Root of kirilow rhodiola (RHODIOLA ROSEA) extract is the extract of root of kirilow rhodiola root, and gas is fragrant and sweet, bitter and puckery flavor, mainly at
Part is rhodioside and Jujubogenin tyrosol, and root of kirilow rhodiola has effective oxidation resistance, the unfavorable work injured by restraint of liberty base
With can effectively resist senescence-factor, be a kind of pure natural antioxidant.
The main component of centella (CENTELLA ASIATICA) extract is Asiaticoside, asiatic acid and hydroxyl product
Oxalic acid is avenged, Gotu Kola P.E can make skin limber up, and help to solve cutis laxa phenomenon (especially postpartum mother), make skin
Smooth flexible, centella can also help to promote the formation of collagen in skin corium, regenerate fibrin, reconnect
Get up, be inherently eliminated wrinkle, achievees the effect that skin compact smooth.Gotu Kola P.E can also help impaired tissue
Healing and consolidation skin, can promote the close connection between epidermis and corium and inhibit the increase of fat cell, prevent skin
There is obesity in skin oedema.
Hydrolysis lupin protein in lupin extract, is able to suppress MMP1,9,3, prevents skin photoage, reinforces true
The junction of epidermis skin strengthens dermal tissue, achievees the effect that skin-tightening.
The present invention is using adipose-derived mescenchymal stem cell excretion body and gadol extract, Gotu Kola P.E and feather fan
These three have effects that the plant extracts for centainly improving wrinkle compounds to beans extract, and the composition is made to have good moisturizing
Moisturizing effect and the effect for improving wrinkle, the composition can obviously increase the moisture content in skin and improve the bullet of skin
Property, keep the effect for going wrinkle and anti-aging of composition more preferable.
Preferably, the weight ratio of adipose-derived the mescenchymal stem cell excretion body and plant extracts are as follows: 1~3:2.
Adipose-derived mescenchymal stem cell excretion body and plant extracts can advantageously promote the suction of plant extracts using said ratio
It receives.It is highly preferred that the weight ratio of adipose-derived the mescenchymal stem cell excretion body and plant extracts are as follows: 1:1.Especially
Said ratio is to promote assimilation effect best, goes wrinkle better effect.
Preferably, the weight ratio of the gadol extract, Gotu Kola P.E and lupin extract are as follows: 1:1~
3:1~3.When three Plant Extracts use said ratio, the inhibitory effect of elastoser and collagenase activity is preferable,
The generation that skin elasticity can be effectively improved, reduce wrinkle.It is highly preferred that the gadol extract, Gotu Kola P.E and
The weight ratio of lupin extract are as follows: 1:2:2.Especially said ratio when, elastoser and collagenase activity
Inhibitory effect is best, and the effect for improving wrinkle is most significant.
Preferably, the adipose-derived mescenchymal stem cell excretion body preparation method the following steps are included:
(1) it disperses adipose tissue in digestive juice, obtains cell suspending liquid；
(2) by cell suspending liquid centrifugal treating, liquid is discarded supernatant, obtains fat stem cell；
(3) fat stem cell is seeded to culture medium and carries out amplification cultivation, centrifugation takes supernatant, is concentrated, and excretion body is added
Extractant obtains the adipose-derived mescenchymal stem cell excretion body.
For the adipose tissue-derived in human or animal, it is Exoquick-TC (SBI) that the excretion body, which extracts reagent, can also
Think that the excretion body of other brands extracts reagent.There is CD9, CD63 mark in the adipose-derived mescenchymal stem cell excretion body
Will.
Preferably, in step (1), the digestive juice is the PBS buffer solution containing collagenase type I；The collagenase type I
Volume is the 0.2~0.4% of adipose tissue volume, and the additional amount of the PBS buffer solution is 4~6 times of adipose tissue volume, is disappeared
The change time is 45~90min.
Preferably, in step (3), the condition of the amplification cultivation are as follows: in incubator, under the conditions of 36~38 DEG C, 5%CO2,
Culture medium is the DMEM culture medium containing 10%FBS, and incubation time is 48~72h.
Preferably, in step (2), the revolving speed of the centrifugation is 1800~2200r/min, and the time is 3~5min.
Preferably, in step (3), after excretion body extractant is added, 8~15h is placed at 3~5 DEG C, is then centrifuged for, takes
Precipitating, i.e., the described adipose-derived mescenchymal stem cell excretion body.
It includes source of human stem cell excretion body for improve wrinkle that the object of the invention is also to provide a kind of comprising described
Composition cosmetics, the cosmetics be aqua, emulsion, cream, gel or pulvis.Except said components clearly require
Outside, in the range of lossless effect of the invention, those skilled in the art can be added according to the dosage form of specific cosmetics or
Use other adjunct ingredients.
Preferably, weight percentage of the adipose-derived mescenchymal stem cell excretion body in the cosmetics is
0.1~1%.
The beneficial effects of the present invention are: it includes source of human stem cell excretion that the present invention provides a kind of for improve wrinkle
The composition and cosmetics of body.Adipose-derived mescenchymal stem cell excretion body is added in composition of the present invention, fat comes
Source mescenchymal stem cell excretion body has the activity for inhibiting elastoser and Collagenase, is effective against aging, improves
The elasticity of skin improves wrinkle of skin, and adipose-derived mescenchymal stem cell excretion body can also promote to a certain extent
Into the absorption of plant extracts, keep the utilization rate of plant extracts in skin higher.
Specific embodiment
Each raw material in embodiment in addition to fatty derived mesenchymal stem cell excretion body is purchased from market.
To better illustrate the object, technical solutions and advantages of the present invention, below in conjunction with specific embodiment to the present invention
It is described further.
Embodiment 1
The present embodiment is a kind of for improving the eye cream of wrinkle, the component comprising following weight percentage: adipose-derived
Mescenchymal stem cell excretion body 0.5%, gadol extract 0.1%, Gotu Kola P.E 0.2%, lupin extract
0.2%, propylene glycol 5%, glycerol 1.6%, stearine 1%, PEG-20 methyl sesquistearate 0.3%, white
Dormant oil 10.2%, dimethyl silicone polymer 2.3%, ammonium acryloyl taurate/VP copolymer 0.5%, carbomer
0.5%, xanthan gum 0.05%, cetanol stearyl alcohol 1.8%, Phenoxyethanol 0.4%, methyl hydroxybenzoate 0.1%, the third rouge of oxybenzene
0.1% and surplus deionized water.
Described in the present embodiment for improve wrinkle eye cream preparation method the following steps are included:
1, by stearine, PEG-20 methyl sesquistearate, White Mineral Oil, dimethyl silicone polymer, hydroxyl
Benzene methyl, the third rouge of oxybenzene sequentially add in oily phase pot, and heating stirring is warming up to 75-85 DEG C；
2, by deionized water, ammonium acryloyl taurate/VP copolymer, cetanol stearyl alcohol, carbomer, xanthan
Glue, propylene glycol, glycerol, Phenoxyethanol, stirring are uniformly mixed, and are heated to 80-90 DEG C, low speed homogeneous keeps material dispersion molten
Solution is complete, and insulated and stirred 10-20 minutes；
3, the mixture that step 1 obtains is added in the mixture that step 2 obtains, is vacuumized, 0.5~0.7MPa, mixed
Homogeneous keeps the temperature 20min, is cooled to 50 DEG C, and propylene glycol is added；40 DEG C are cooled to, addition will be outside adipose-derived mescenchymal stem cell
Body, gadol extract, Gotu Kola P.E, lupin extract are secreted, is stirred evenly, is obtained described for improving the eye of wrinkle
Frost.
The preparation method of adipose-derived mescenchymal stem cell excretion body described in the present embodiment the following steps are included:
(1) it disperses human fat tissue in the PBS buffer solution digestive juice containing collagenase type I, the collagenase type I
Volume is the 0.3% of adipose tissue volume, and the additional amount of the PBS buffer solution is 5 times of adipose tissue volume, and digestion time is
60min obtains cell suspending liquid；
(2) cell suspending liquid is discarded supernatant liquid, obtains fat stem cell in 2000r/min centrifugal treating 4min；
(3) fat stem cell is seeded to culture medium and carries out amplification cultivation, the condition of the amplification cultivation are as follows: in incubator
It is interior, under the conditions of 36~38 DEG C, 5%CO2, culture medium is the DMEM culture medium containing 10%FBS, and incubation time is 72h centrifugation, is taken
Clear liquid is concentrated into 50% or more volume-diminished, is added excretion body extractant Exoquick-TC (SBI), and excretion body extractant adds
Entering amount is the supernatant after concentration 50% to get the adipose-derived mescenchymal stem cell excretion body.
Embodiment 2
One, elastase inhibitory activity
Substrate N- succinyl-alanine-alanine-alanine-p- nitro is acted on by pancreatic elastase by measuring
The amount of 4- nitroaniline (4-nitoanilide) peptide fragment that aniline generates utilizes the work of spectrophotometry assessment elastoser
Property.N- succinyl-alanine-alanine-alanine-p-NA Sigma) and it is other for test substance at 37 DEG C
The reaction of porcine pancreatic elastase initial is added in lower incubation.Measurement wavelength be 410nm place absorbance come measure be released dissociate
The amount of 4- nitroaniline.The Tris-HCl buffer that the pH of 0.2M is 8.0 is used as negative control, and test group and test result are shown in Table
1。
1 elastase activity test result of table
It can be seen that adipose-derived from the percentage that 1 elastase enzyme suppression percentage of table is opposite negative control
Mesenchymal stem cells excretion body has dose-dependant formula inhibitory effect to elastase activity, and plant extracts is to elastin laminin
Enzymatic activity has certain inhibitory effect.
Two, Collagenase inhibition analysis
Collagenase (enzyme EC 3.4.24.3,5 μ g) individually with adipose-derived mescenchymal stem cell excretion body (1 μ g/mL
It is added to together in the Tris buffer (pH=7.4) of the 0.1M containing PZ peptide (0.5mg) respectively with 10 μ g/mL), makes total volume
For 1.7ml, PZ peptide is the substrate of Collagenase.Mixture incubates 30min in 37 DEG C of water-baths, and the 25mM of 1mL is then added
Citric acid solution terminate enzyme reaction.After the ethyl acetate mixing of 5mL is added, it is in wavelength using ultraviolet specrophotometer
The absorbance of organic layer is measured at 320nm.Collagenase inhibition is calculated by following formula:
Collagenase suppression percentage=[(Ac-As)/Ac] × 100, wherein Ac expression (contains Collagenase
The absorbance of control of the absorbance-of control without Collagenase), As indicates the (extinction of the sample containing Collagenase
The absorbance of sample of the degree-without Collagenase).
Compared with the control, the Collagenase of 0.1% and 1.0% adipose-derived mescenchymal stem cell excretion body inhibits hundred
Point than being respectively 88.5% and 62.8%, collagen enzyme inhibition activity is shown, and (mass concentration is plant extracts
0.1% gadol extract, 0.2% Gotu Kola P.E, 0.2% lupin extract mixture) Collagenase
Suppression percentage is 84.0%, shows that plant extracts has certain inhibitory effect to Collagenase.
Gadol extract, Gotu Kola P.E and lupin identical to concentration but different ratio using same method
Extract has carried out elastoser and collagenase activity inhibits test, and test result shows gadol extract, accumulated snow
The inhibitory effect of two kinds of enzymes is preferable when the weight ratio of careless extract and lupin extract is 1:1~3:1~3, especially
When the weight ratio of gadol extract, Gotu Kola P.E and lupin extract is 1:2:2, inhibitory effect is best.
Three, to verify effect of the composition of the present invention in terms of going wrinkle, 30 healths are selected, without skin care item mistake
The volunteer of quick history, volunteer were the age between 40~55 years old, were randomly divided into 3 groups, men and women is unlimited, every group of 10 people, respectively
Test group 10~12.
The skin care item that test group 10~12 uses respectively are as follows:
Test group 10: eye cream described in embodiment 1；
Test group 11: gadol extract 0.2%, Gotu Kola P.E 0.4%, lupin extract 0.4%, propylene glycol
5%, glycerol 1.6%, stearine 1%, PEG-20 methyl sesquistearate 0.3%, White Mineral Oil 10.2%, poly-
Dimethyl siloxane 2.3%, ammonium acryloyl taurate/VP copolymer 0.5%, carbomer 0.5%, xanthan gum
0.05%, cetanol stearyl alcohol 1.8%, Phenoxyethanol 0.4%, methyl hydroxybenzoate 0.1%, the third rouge of oxybenzene 0.1% and surplus are gone
Ionized water.
Test group 12: adipose-derived mescenchymal stem cell excretion body 1%, propylene glycol 5%, glycerol 1.6%, glycerol stearate
Ester 1%, PEG-20 methyl sesquistearate 0.3%, White Mineral Oil 10.2%, dimethyl silicone polymer 2.3%, acryloyl
Dimethyltaurine ammonium/VP copolymer 0.5%, carbomer 0.5%, xanthan gum 0.05%, cetanol stearyl alcohol 1.8%, benzene oxygen
The deionized water of ethyl alcohol 0.4%, methyl hydroxybenzoate 0.1%, the third rouge of oxybenzene 0.1% and surplus.
After the daily morning and evening face cleaning of 3 groups of volunteers, using the skin care item, application method is to take to be applied to eye circumference in right amount, gently
It gently smears by fully absorbing.Regular follow-up investigation is carried out to volunteer, using skinanalysis apparatus (Derma-Expert MC760)
Skin elasticity test is carried out to skin, test in every 7 days is primary, and test position is eye circumference, specially outside canthus.Record is each respectively
Mean value (the D of skin parameters when group volunteer's on-test0), the mean value (D of skin parameters when testing the 7th day7), Yi Jishi
Mean value (the D of skin parameters when testing the 14th day14), and calculate and test the 7th day and items skin parameters mean value when on-test
Difference (D7-D0) and test the 14th day with test the 7th day when items skin parameters mean value difference (D14-D7).When volunteer's items
When parameter difference >=5, being considered as skin improves crowd.Skin improvement rate=each group skin improvement number/each group total number of persons ×
100%.Moisture of skin test condition is 20 DEG C of temperature and relative humidity is 40~60%.Skin elasticity test result is shown in Table 2.
2 skin elasticity test result of table
| Group | D7-D0 | D14-D7 | Skin improvement rate (%) |
| Test group 10 | 11.46 | 10.25 | 100 |
| Test group 11 | 5.67 | 5.63 | 60 |
| Test group 12 | 5.56 | 6.04 | 80 |
From table 2 it can be seen that adipose-derived mescenchymal stem cell excretion body, gadol extract, Gotu Kola P.E,
Under total addition level unanimous circumstances of the lupin extract in eye cream, using eye cream described in test group 10 improvement more
Significantly, illustrate that adipose-derived mescenchymal stem cell excretion body may be by promoting absorption of the skin to plant extracts, to send out
That waves synergy goes wrinkle effect.
Using the above method to adipose-derived mescenchymal stem cell excretion body, gadol extract, Gotu Kola P.E and
Lupin extract is identical in total addition level, but is tested when different ratio, research shows that working as adipose-derived mesenchyma
When the weight ratio of stem cell excretion body and plant extracts is 1~3:2, there is preferable improvement to wrinkle, and fat comes
The weight ratio of source mescenchymal stem cell excretion body and plant extracts is best when being 1:1.And when adipose-derived mesenchyma is dry
When the additive amount of cell excretion body is 0.1~1%, preferable facilitation effect can be realized.
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than protects to the present invention
The limitation of range is protected, although the invention is described in detail with reference to the preferred embodiments, those skilled in the art should
Understand, it can be with modification or equivalent replacement of the technical solution of the present invention are made, without departing from the essence of technical solution of the present invention
And range.
Claims (10)
1. a kind of for improving the composition comprising source of human stem cell excretion body of wrinkle, which is characterized in that comprising adipose-derived
Mescenchymal stem cell excretion body and plant extracts.
2. as described in claim 1 for improving the composition comprising source of human stem cell excretion body of wrinkle, which is characterized in that institute
Stating plant extracts includes gadol extract, Gotu Kola P.E and lupin extract.
3. as described in claim 1 for improving the composition comprising source of human stem cell excretion body of wrinkle, which is characterized in that institute
State the weight ratio of adipose-derived mescenchymal stem cell excretion body and plant extracts are as follows: 1~3:2；Preferably, the fat comes
The weight ratio of source mescenchymal stem cell excretion body and plant extracts are as follows: 1:1.
4. as described in claim 1 for improving the composition comprising source of human stem cell excretion body of wrinkle, which is characterized in that institute
State the weight ratio of gadol extract, Gotu Kola P.E and lupin extract are as follows: 1:1~3:1~3；Preferably, described
The weight ratio of gadol extract, Gotu Kola P.E and lupin extract are as follows: 1:2:2.
5. as described in claim 1 for improving the composition comprising source of human stem cell excretion body of wrinkle, which is characterized in that institute
State the preparation method of adipose-derived mescenchymal stem cell excretion body the following steps are included:
(1) it disperses adipose tissue in digestive juice, obtains cell suspending liquid；
(2) by cell suspending liquid centrifugal treating, liquid is discarded supernatant, obtains fat stem cell；
(3) fat stem cell is seeded to culture medium and carries out amplification cultivation, centrifugation takes supernatant, is concentrated, and excretion body is added and extracts
Agent obtains the adipose-derived mescenchymal stem cell excretion body.
6. as claimed in claim 5 for improving the composition comprising source of human stem cell excretion body of wrinkle, which is characterized in that step
Suddenly in (1), the digestive juice is the PBS buffer solution containing collagenase type I；The volume of the collagenase type I is adipose tissue body
Long-pending 0.2~0.4%, the additional amount of the PBS buffer solution are 4~6 times of adipose tissue volume, digestion time is 45~
90min。
7. as claimed in claim 5 for improving the composition comprising source of human stem cell excretion body of wrinkle, which is characterized in that step
Suddenly in (3), the condition of the amplification cultivation are as follows: in incubator, under the conditions of 36~38 DEG C, 5%CO2, culture medium is containing 10%FBS
DMEM culture medium, incubation time be 48~72h.
8. as claimed in claim 2 for improving the composition comprising source of human stem cell excretion body of wrinkle, which is characterized in that step
Suddenly in (3), after excretion body extractant is added, 8~15h is placed at 3~5 DEG C, is then centrifuged for, take precipitating, i.e., the described fat comes
Source mescenchymal stem cell excretion body.
9. it is a kind of comprising as described in any one of claim 1~8 for improve wrinkle comprising source of human stem cell excretion body
The cosmetics of composition, which is characterized in that the cosmetics are aqua, emulsion, cream, gel or pulvis.
10. cosmetics as claimed in claim 9, which is characterized in that the adipose-derived mescenchymal stem cell excretion body is described
Weight percentage in cosmetics is 0.1~1%.
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| CN111821252A (en) * | 2020-08-12 | 2020-10-27 | 辽宁盛京干细胞科技有限公司 | Anti-wrinkle essence containing adipose-derived stem cell extract and preparation method thereof |
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| CN115252531A (en) * | 2022-08-11 | 2022-11-01 | 睿博斯(北京)生物科技有限公司 | Anti-wrinkle essence and preparation method thereof |
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| CN111821252A (en) * | 2020-08-12 | 2020-10-27 | 辽宁盛京干细胞科技有限公司 | Anti-wrinkle essence containing adipose-derived stem cell extract and preparation method thereof |
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| CN114796089B (en) * | 2022-05-26 | 2024-05-14 | 格莱康美生物医学技术(北京)有限公司 | Application of umbilical cord mesenchymal stem cell exosome combined rhodiola rosea polypeptide in maintaining beauty and keeping young |
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Application publication date: 20190823 |