CN108837142A - A kind of stem cell excretion body and its preparation method and application for repairing skin - Google Patents
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Abstract
The present invention provide it is a kind of can repair stem cell excretion body of skin and its preparation method and application, wherein preparation method is decapeptide -12 to be transduceed in mescenchymal stem cell excretion body to get to the stem cell excretion body that can repair skin.The stem cell excretion body for repairing skin prepared through the invention, it finds for mescenchymal stem cell excretion body and decapeptide -12 to be combined for the first time, utilize the load capacity and lower immunogenicity of excretion body cell, it so that decapeptide -12 is reached melanocyte and then inhibit tyrosinase activity, inflammatory reaction can also can be adjusted, the regeneration of skin relevant cell is promoted.The stem cell excretion body prepared by the present invention for repairing skin, which is applied to the skin after tanning severely, can achieve the effect that eliminate erythema, oedema, it more can be reduced the generation of melanin and then reduce the formation of blackspot after pigment deposition and sunburn, it can be used in the fields such as cosmetics and treatment skin disease drug, with significant effect and there is important application value.
Description
Technical field
The present invention relates to biological cell technical field, in particular to a kind of stem cell excretion body and its system that can repair skin
Preparation Method and application.
Background technique
Sunlight is made of the light wave of different wave length, wherein ultraviolet B radiation (UVB) can reach the substrate of epidermis
Layer, it is strong epidermal keratinocyte to be caused downright bad according to solarization, and the inflammatory mediators such as histamine, serotonin, kassinin kinin are discharged, and then cause
Dermal vascular expansion and tissue edema, followed by melanocyte accelerate synthetic dyestuff, deepen skin pigment.Skin sunburn after solarization
Be mainly shown as that erythema, oedema or blister occurs in local skin, more after there are pigmentation spots, and with burn, feeling of pain.
Studies have shown that UVB can not only coup injury cell, and the microRNA meeting being damaged that damaged cell is discharged
The signal tanned severely as skin releases, and excites neighbouring healthy cell, to cause inflammation (Bernard et
al.,2012).Excretion body secreted by skin keratinocytes after equally receiving UVB irradiation, will induce melanocyte
(melanocyte) pigment is generated, and then shows skin pigment after shining and deepens (Lo Cicero et al., 2015).It is more tight
Weight, the inflammatory reaction after ultraviolet irradiation can promote the blood vessel of melanoma to orient and transfer (Bald et al., 2014),
Further increase the transfer of melanoma.In short, skin sunburn will induce inflammatory reaction, erythema, oedema are directly translated into, and
Melanocyte increases, pigment synthesis exacerbation leads to problems such as skin color deepen and formed pigment deposition spot.
In existing substance having sun-screening function, as (English name Decapeptide-12, trade name Lumixyl or dew are bright for decapeptide -12
This), amino acid sequence YRSRKYSSWY is a kind of new and effective and extremely low cytotoxicity tyrosinase inhibitor, makees
It is added in cosmetics the generation that can effectively inhibit melanin for cosmetic material.However, existing substance having sun-screening function is only to suppression
Melanin production processed has a certain effect, and is but difficult to play good repair to the skin after having tanned severely.
Summary of the invention
To solve the problems, such as to mention in above-mentioned background technique, the present invention provides a kind of stem cell excretion body that can repair skin
Preparation method, method is：Decapeptide -12 is transduceed in mescenchymal stem cell excretion body to get to can repair the dry thin of skin
It is extracellular to secrete body.
Further, the mescenchymal stem cell excretion body includes umbilical cord mesenchymal stem cells, placenta mesenchyma stem cell
And fat mesenchymal stem cell.
Further, decapeptide -12 is transduceed and is included the following steps in mescenchymal stem cell excretion body：
Step a, mescenchymal stem cell excretion body is dissolved in the PBS buffer solution containing trehalose, forms mixed liquor M1；
Step b, M1 and decapeptide -12 are separately added into electroporation ware, are put into electroporation apparatus, carry out electroporation transduction；
Step c, after transduction using PBS buffer solution wash, with containing trehalose PBS buffer solution be resuspended precipitating to get
To the stem cell excretion body that can repair skin.
Further, in step b, when carrying out electroporation transduction, voltage 400V, capacitor is 250 μ F.
Further, it after precipitating being resuspended with PBS buffer solution, after -80 DEG C or less freeze, is transferred in vacuum freeze-drying storehouse
Freeze-drying, after obtaining the freeze-dried powder for the stem cell excretion body that can repair skin, is saved with spare.
Further, the vacuum degree in the vacuum freeze-drying storehouse is 10Pa, and temperature is -45 DEG C, cooling time 12h-
48h。
The present invention also provides the stem cell excretion bodies that skin is repaired prepared by a kind of as above any preparation method.
The present invention also provides the stem cell excretion bodies that skin is repaired prepared by a kind of as above any preparation method
Preparing the application on cosmetics.
The present invention also provides the stem cell excretion bodies that skin is repaired prepared by a kind of as above any preparation method
Application on preparation treatment skin disease drug.
In the stem cell excretion body provided by the invention for repairing skin, mescenchymal stem cell excretion body not only has adjusting
The effect of inflammatory reaction, the more regeneration of promotion skin relevant cell and transfer ability (Wu, Zhang, Shi, Qian , &Xu,
2018), and Decapeptide-12 be a kind of new and effective and extremely low cytotoxicity tyrosinase inhibitor (Abu Ubeid,
Zhao,Wang,&Hantash,2009)。
The stem cell excretion body for repairing skin prepared through the invention, finds for the first time by mescenchymal stem cell excretion body
It is combined with decapeptide -12, using the load capacity and lower immunogenicity of excretion body cell, can not only make decapeptide -12
It reaches melanocyte and then inhibits tyrosinase activity, additionally it is possible to inflammatory reaction can be adjusted, the regeneration of skin relevant cell is promoted,
Applied to the skin after sunburn can reach eliminate erythema, oedema, more can be reduced melanin generation so that reduce pigment deposition and
The formation of blackspot after sunburn can use in the fields such as cosmetics and treatment skin disease drug, have significant effect and have
There is important application value.
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, it should be apparent that, the accompanying drawings in the following description is this hair
Bright some embodiments for those of ordinary skill in the art without any creative labor, can be with
Other attached drawings are obtained according to these attached drawings.
Fig. 1 is the test result of the UV absorption test of the sample progress 280nm of the embodiment of the present invention and comparative example preparation
Figure；
Fig. 2 is the partial size test result figure using the NTA excretion body obtained to different modes；
Fig. 3 is the particle concentration test result figure using the NTA excretion body obtained to different modes.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is
A part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art
Every other embodiment obtained without creative efforts, shall fall within the protection scope of the present invention.
The present invention provides EXPERIMENTAL EXAMPLE in detail below, includes the following steps：
One, mescenchymal stem cell culture：
The mescenchymal stem cell of use is dry comprising umbilical cord mesenchymal stem cells, placenta mesenchyma stem cell, fat mesenchymal
Cell etc., the present invention in using umbilical cord mesenchymal stem cells as experimental subjects.
Step a, the umbilical cord mesenchymal stem cells cryopreservation tube in P3 generation is taken out from liquid nitrogen container, is immediately placed in 37 DEG C of water-baths
It thaws 2 minutes, to melt completely, freezes tube wall with 75% alcohol wipe and be placed in Biohazard Safety Equipment；
Step b, the umbilical cord mesenchymal stem cells suspension after recovery is transferred in T75 culture bottle, by what is incubated
30ml cell culture medium (+100 μ g/ml streptomysin of DMEM/F12+10%FBS+2mM l-Glutathione+100U/ml penicillin：
Basal medium can use α-MEM or L-DMEM culture medium) T75 is added, and (concentration of stem cells is 4 × 106/ ml), at 37 DEG C, 5%
CO2Incubator is incubated overnight；Second day adherent situation of observation cell and cellular morphology, it is rear to change liquid with the new culture medium incubated.
Step c, when 70%-85% is merged, secondary culture can be started, concrete operations are：It is cleaned using PBS buffer solution
Twice of the cell of adherent growth is digested to cell with 0.25% pancreatin and takes off wall, is added in fresh culture medium and pancreatin, according to
1:3 ratio carries out cell secondary culture (20% fusion is advisable after inoculation).
Step d, it changes liquid within 3 days or so, and observes the fusion situation of cell；To 70%-80% cell fusion, it is replaced with excretion
Body collects culture medium, and (DMEM/F12+10% is without+100 μ g/ml chain of excretion body FBS+2mM l-Glutathione+100U/ml penicillin
+ 7 μm of multipotency rhzomorphs of mycin：Basal medium can use α-MEM or L-DMEM culture medium), it is further cultured for 48-72 hours, and observe
Cell fusion situation and form.
Step e, addition is collected culture medium and is further cultured for 48-72 hours, microscopy results, when cell fusion reaches 90%
When above, that is, obtain cell culture fluid.
Two, the separation of mescenchymal stem cell excretion body
The cell culture supernatant liquid of above-mentioned acquisition is placed in centrifuge tube, under the conditions of 4 DEG C, 5min is centrifuged with 500g
Afterwards, centrifuge tube supernatant is regathered；
Under the conditions of 4 DEG C, after 2000g centrifugation 30min removal cell fragment and protein polymer, supernatant is collected again
Liquid, and cross 0.22 μm of filter membrane, that is, obtain the excretion body crude extract of removal cell, cell fragment etc.；
By excretion body crude extract, under the conditions of 4 DEG C, 1h is centrifuged with 10000g, big apoptotic bodies are removed, in 4 DEG C of items
100000g is centrifuged 2h under part, is resuspended and is precipitated with 200 μ l PBS buffer solutions, i.e., isolated mescenchymal stem cell excretion body, will
Mescenchymal stem cell excretion body after separation freezes at -80 DEG C, with spare；The use of mescenchymal stem cell excretion body will be prepared
NTA measures the partial size and concentration of excretion body, and test result is as follows shown in table：
Table 1
| Particle concentration (109/ml) | Standard deviation | NTA partial size (nm) | Standard deviation |
| 2.7 | 0.5 | 93 | 5 |
As seen from the above table, the mescenchymal stem cell excretion bulk concentration and partial size and document report extracted by supercentrifugation
Road unanimously to get to mescenchymal stem cell excretion body can be applied to subsequent experiment.
Three, the preparation of the stem cell excretion body of skin can be repaired
By 2 × 109The excretion body of particles is dissolved in the PBS buffering that 400 μ l contain 50mM trehalose (Terahorse)
(4 DEG C) formation mixed liquor M1 in liquid；
By the Decapeptide-12 (Lumixyl of 1mgTM,Envy Medical,Inc.Westlake Village,USA)
And mixed liquor M1 is separately added into 0.4cm (750 μ l of volume) electroporation ware, electroporation apparatus is put into, in voltage 400V, 250 μ of capacitor
Electroporation transduction is carried out under the conditions of F；
(4 DEG C, 100000g is centrifuged 2h) are washed twice after transduction with PBS buffer solution, with 200 μ l with 200 μ l seaweed containing 30mM
Precipitating is resuspended to get to the stem cell excretion body that can repair skin in the PBS buffer solution of sugar；
The obtained stem cell excretion body for repairing skin is frozen in -80 DEG C, it is spare.
Due to containing 1 trp residue on Decapeptide-12,3 tyrosine (Tyr) residues, therefore can will be upper
State embodiment preparation stem cell excretion by measurement electroporation import after 280nm ultraviolet absorption peak come identify import whether at
Function；Meanwhile the following control of setting：
Blank control：400 μ l contain the PBS buffer solution of 50mM trehalose (Terahorse)；
Comparative example a, Decapeotide-12 is not added as control：2×1010The excretion body of particles is dissolved in 400 μ l
PBS buffer solution containing 50mM trehalose (Terahorse)；
Comparative example b, the transduction control of non-electroporation：The Decapeptide-12 of 1mg, 2 × 109The excretion body of particles
It is dissolved in the PBS buffer solution that 400 μ l contain 50mM trehalose (Terahorse).
A group sample treatment process is consistent with experimental group in the above comparative example, and b group is other in addition to not carrying out electroporation transduction
It is completely the same with experimental group process.
The sample of Example and comparative example preparation respectively, measurement is in the ultraviolet inspiration market condition of 280nm, and test result is such as
Shown in Fig. 1, as shown in Figure 1, after electroporated transduction and elution is gone after not importeding into the Decapeptide-12 in excretion body, sample
Ultraviolet absorption value of the product in 280nm reaches 0.19, and the ultraviolet absorption value for comparing two experiment contrasts proves to turn by electroporation
Decapeptide-12 is led effectively to imported into excretion body.
By the obtained stem cell excretion body for repairing skin after -80 DEG C freeze 12h, it is transferred to vacuum freeze-drying storehouse jelly
Dry, the vacuum degree of freeze-drying is 10Pa, and temperature is -45 DEG C hereinafter, cooling time is 12h-48h；It is dry that mesenchyma is obtained after freeze-drying
Cell excretion body freeze-dried powder is kept in dark place under the conditions of 4 DEG C after product is made；Skin can be repaired by made from as the above method
Stem cell excretion body prepare at freeze-dried powder, the storage and transport of product can be conducive to.
Mescenchymal stem cell excretion body (the i.e. unmodified mesenchyma that the separation method provided through the invention is obtained
Stem cell excretion body), the stem cell excretion body (i.e. electroporation transduction after) for repairing skin of preparation of the embodiment of the present invention and
Outside the stem cell that the stem cell excretion body freeze-dried powder for repairing skin of preparation is redissolved after storing one month with PBS buffer solution
Body (i.e. freeze-drying is redissolved) to be secreted, partial size and concentration are detected using NTA, the results of comparison of detection is as shown in Figures 2 and 3, as seen from the figure,
It is redissolved after electroporated transduction and freeze-drying, the partial size and concentration of excretion body will not give birth to it in acceptable range
Object function effectively influences greatly.
The stem cell excretion body for repairing skin that the present invention is prepared carries out following validity test：
Personage after beauty parlor selects 20 skin sunburns is divided into two groups, and one group is repaired using provided by the invention
The stem cell excretion body freeze-dried powder of skin is treated after redissolving (10 after freeze-dried powder redissolution9Particles/ml is applied to and shines injury
And other are exposed at daylight), it is treated using conventional sunburn treatment method for another group and (takes Histamine agents object cyproheptadine
2mg three times a day takes orally, while taking anti-oxidation medicine vitamin C and composite microbial element B three times a day takes orally).Complete one
After the treatment of a course for the treatment of (15 days), assay is as a result, test result is as follows shown in table, wherein table 1 is provides using the present invention
The stem cell excretion body freeze-dried powder for repairing skin redissolve after treated as a result, table 2 for using conventional sunburn treatment side
The result that method is treated；
Wherein, in inflammation inhibitory effect：
Effective evaluation criterion is in -12 hours, and oedema disappears, red and swollen to disappear with pain；
Good evaluation criterion is oedema decline in -24 hours, red and swollen to disappear with pain；
General evaluation criterion is oedema decline in -48 hours, red and swollen to disappear with pain；
Invalid evaluation criterion is in -72 hours, and oedema does not decline, and redness does not disappear with pain.
Anti-melanin is formed in effect：
Effective evaluation criterion is-after a course of therapy, take pictures with treat before photo comparison, skin has no melanin
Deposition.
Good evaluation criterion is-after a course of therapy, take pictures with treat before photo comparison, skin is slightly shown in melanin
Deposition.
General evaluation criterion is-after a course of therapy, take pictures with treat before photo comparison, skin has to a certain degree
Melanin deposition, but subject's self-assessment is effective.
Invalid evaluation criterion is-after a course of therapy, take pictures with treat before photo comparison, skin is shown in obvious black
Element deposition.
Table 1
Table 2
The stem cell excretion body freeze-dried powder provided by the invention for repairing skin is redissolving it can be seen from Tables 1 and 2
Afterwards, the skin of sunburn is treated, can not only can has the function that eliminate erythema, oedema, more can be reduced the life of melanin
At the formation of blackspot after reducing pigment deposition in turn and tanning severely, there is significant repair to the skin after sunburn.
The present invention also provides a kind of stem cell excretion bodies for repairing skin provided using this technology to prepare cosmetics
With the application on preparation treatment skin disease drug, valence is applied to reparation skin with positive beneficial effect and with important
Value.
Wherein, reference paper involved in the invention patent is as follows：
Abu Ubeid,A.,Zhao,L.,Wang,Y.,&Hantash,B.M.(2009).Short-sequence
oligopeptides with inhibitory activity against mushroom and human
tyrosinase.J Invest Dermatol,129(9),2242-2249.doi:10.1038/jid.2009.124
Bald,T.,Quast,T.,Landsberg,J.,Rogava,M.,Glodde,N.,Lopez-Ramos,D.,
Tuting,T.(2014).Ultraviolet-radiation-induced inflammation promotes
angiotropism and metastasis in melanoma.Nature,507(7490),109-113.doi:10.1038/
nature13111
Bernard,J.J.,Cowing-Zitron,C.,Nakatsuji,T.,Muehleisen,B.,Muto,J.,
Borkowski,A.W.,...Gallo,R.L.(2012).Ultraviolet radiation damages self
noncoding RNA and is detected by TLR3.Nat Med,18(8),1286-1290.doi:10.1038/
nm.2861
Lo Cicero,A.,Delevoye,C.,Gilles-Marsens,F.,Loew,D.,Dingli,F.,Guere,
C.,...Raposo,G.(2015).Exosomes released by keratinocytes modulate melanocyte
pigmentation.Nat Commun,6,7506.doi:10.1038/ncomms8506
Wu,P.,Zhang,B.,Shi,H.,Qian,H.,&Xu,W.(2018).MSC-exosome:A novel cell-
free therapy for cutaneous regeneration.Cytotherapy,20(3),291-301.doi:
10.1016/j.jcyt.2017.11.002
Finally it should be noted that：The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；It is real
It applies and is related to reagent concentration and other parameters in example for a kind of feasible pattern under present inventive concept, be not restricted to this reality
Apply related experiment parameter defined in example；Under present inventive concept, Related Technical Issues are able to solve, reach the technology of the present invention
The related experiment parameter of effect is within the protection scope of the present invention；Although having been carried out in detail referring to foregoing embodiments to the present invention
Thin explanation, those skilled in the art should understand that：It still can be to technical side documented by foregoing embodiments
Case is modified, or equivalent substitution of some or all of the technical features；And these are modified or replaceed, not
The essence of corresponding technical solution is set to depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (9)
1. the preparation method that one kind can repair the stem cell excretion body of skin, it is characterised in that：
Decapeptide -12 is transduceed in mescenchymal stem cell excretion body to get to the stem cell excretion body that can repair skin.
2. the preparation method of the stem cell excretion body according to claim 1 for repairing skin, it is characterised in that：Described
Mesenchymal stem cells excretion body includes one in umbilical cord mesenchymal stem cells, placenta mesenchyma stem cell and fat mesenchymal stem cell
Kind.
3. the preparation method of the stem cell excretion body according to claim 1 for repairing skin, which is characterized in that by ten
Peptide -12, which is transduceed, to be included the following steps in mescenchymal stem cell excretion body：
Step a, mescenchymal stem cell excretion body is dissolved in the PBS buffer solution containing trehalose, forms mixed liquor M1；
Step b, M1 and decapeptide -12 are separately added into electroporation ware, electroporation transduction is carried out using electroporation apparatus；
Step c, it is washed after transduction using PBS buffer solution, precipitating is resuspended to get to can with the PBS buffer solution containing trehalose
Repair the stem cell excretion body of skin.
4. the preparation method of the stem cell excretion body according to claim 3 for repairing skin, it is characterised in that：Step b
In, when carrying out electroporation transduction, voltage 400V, capacitor is 250 μ F.
5. the preparation method of the stem cell excretion body according to claim 3 for repairing skin, it is characterised in that：Use PBS
After precipitating is resuspended in buffer solution, after -80 DEG C or less freeze, it is transferred in vacuum freeze-drying storehouse and is lyophilized, obtain that skin can be repaired
After the freeze-dried powder of stem cell excretion body, saved with spare.
6. the preparation method of the stem cell excretion body according to claim 5 for repairing skin, it is characterised in that：It is described true
The vacuum degree of empty dry storehouse is 10Pa, and temperature is -45 DEG C, cooling time 12h-48h.
7. repairing the stem cell excretion body of skin prepared by a kind of any one of -6 preparation methods according to claim 1.
8. the stem cell excretion body for repairing skin prepared by a kind of any one of -6 preparation methods according to claim 1 exists
Prepare the application on cosmetics.
9. the stem cell excretion body for repairing skin prepared by a kind of any one of -6 preparation methods according to claim 1 exists
Application on preparation treatment skin disease drug.
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109355259A (en) * | 2018-11-23 | 2019-02-19 | 北京太东生物科技有限公司 | A kind of umbilical cord mesenchymal stem cells excretion body culture and separation method |
| CN110279893A (en) * | 2019-08-13 | 2019-09-27 | 成都清科生物科技有限公司 | A kind of excretion body freeze-dried powder and preparation method thereof and the preparation comprising the excretion body freeze-dried powder |
| CN110538197A (en) * | 2019-09-27 | 2019-12-06 | 北京臻惠康生物科技有限公司 | Application of exosome in medicine for treating alopecia |
| CN111110699A (en) * | 2020-04-01 | 2020-05-08 | 北京岳昊科技发展有限公司 | Application of combination of polypeptide conjugate and epidermal stem cell exosome in medicines and cosmetics |
| CN111297900A (en) * | 2020-05-13 | 2020-06-19 | 优赛生命科技有限公司 | Composition rich in adipose-derived stem cell exosomes and application thereof |
| CN111643527A (en) * | 2020-07-22 | 2020-09-11 | 北京岳昊科技发展有限公司 | Application of transgenic stem cell exosome in preparation of medicine or whitening cosmetics |
| CN112386687A (en) * | 2020-12-09 | 2021-02-23 | 北京欣颂生物科技有限公司 | Stem cell exosome and application thereof in medicines and cosmetics |
| CN112472621A (en) * | 2020-12-09 | 2021-03-12 | 北京欣颂生物科技有限公司 | Preparation method of stem cell exosome and medicine or cosmetic prepared by stem cell exosome |
| CN113248573A (en) * | 2021-05-18 | 2021-08-13 | 北京戴域生物技术有限公司 | Application of active peptide and mesenchymal stem cell exosome for improving skin physiological characteristics in medicines or cosmetics |
| CN113307851A (en) * | 2021-06-11 | 2021-08-27 | 北京戴域生物技术有限公司 | Application of active peptide and mesenchymal stem cell exosome for improving skin physiological characteristics in medicines or cosmetics |
| CN113755445A (en) * | 2021-07-26 | 2021-12-07 | 蓝海细胞(北京)生物科技有限公司 | Preparation method of cell exosome for skin repair |
| CN114159374A (en) * | 2021-12-10 | 2022-03-11 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method, product and application of skin anti-aging mesenchymal stem cell exosome composition |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110091530A1 (en) * | 2007-06-27 | 2011-04-21 | The Board Of Trustees Of The Leland Stanford Junior University | Oligopeptide tyrosinase inhibitors and uses thereof |
| CN104490931A (en) * | 2014-12-17 | 2015-04-08 | 江苏大学 | Application of exosome secreted by human umbilical cord mesenchymal stem cells to treating skin injury |
| CN105935343A (en) * | 2016-05-30 | 2016-09-14 | 哈尔滨源茂达生物技术有限公司 | Whitening and spot removing polypeptide essence and preparation method thereof |
| CN107007541A (en) * | 2017-05-23 | 2017-08-04 | 北京希诺赛尔健康科技推广有限公司 | Application of the excretion body in skin-whitening preparation |
-
2018
- 2018-07-20 CN CN201810799770.5A patent/CN108837142A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110091530A1 (en) * | 2007-06-27 | 2011-04-21 | The Board Of Trustees Of The Leland Stanford Junior University | Oligopeptide tyrosinase inhibitors and uses thereof |
| CN104490931A (en) * | 2014-12-17 | 2015-04-08 | 江苏大学 | Application of exosome secreted by human umbilical cord mesenchymal stem cells to treating skin injury |
| CN105935343A (en) * | 2016-05-30 | 2016-09-14 | 哈尔滨源茂达生物技术有限公司 | Whitening and spot removing polypeptide essence and preparation method thereof |
| CN107007541A (en) * | 2017-05-23 | 2017-08-04 | 北京希诺赛尔健康科技推广有限公司 | Application of the excretion body in skin-whitening preparation |
Non-Patent Citations (4)
| Title |
|---|
| STEVE PARKER: "《人体》", 31 January 2014, 上海科学技术出版社 * |
| 刘丽红 等: ""十肽-12治疗黄褐斑临床疗效观察"", 《实用皮肤病学杂志》 * |
| 李思迪 等: ""外泌体: 为高效药物投递策略提供天然的内源性纳米载体"", 《化学进展》 * |
| 魏晓晶 等: ""外泌体功能与临床应用研究进展"", 《中国医药导报》 * |
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| CN109355259A (en) * | 2018-11-23 | 2019-02-19 | 北京太东生物科技有限公司 | A kind of umbilical cord mesenchymal stem cells excretion body culture and separation method |
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| CN112386687A (en) * | 2020-12-09 | 2021-02-23 | 北京欣颂生物科技有限公司 | Stem cell exosome and application thereof in medicines and cosmetics |
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| CN113248573A (en) * | 2021-05-18 | 2021-08-13 | 北京戴域生物技术有限公司 | Application of active peptide and mesenchymal stem cell exosome for improving skin physiological characteristics in medicines or cosmetics |
| CN113248573B (en) * | 2021-05-18 | 2022-03-22 | 深圳市旷逸生物科技有限公司 | Application of active peptide and mesenchymal stem cell exosome for improving skin physiological characteristics in medicines or cosmetics |
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| CN113755445A (en) * | 2021-07-26 | 2021-12-07 | 蓝海细胞(北京)生物科技有限公司 | Preparation method of cell exosome for skin repair |
| CN114159374A (en) * | 2021-12-10 | 2022-03-11 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method, product and application of skin anti-aging mesenchymal stem cell exosome composition |
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