CN109432127A - Application of the mescenchymal stem cell excretion body in the pharmaceutical preparation that preparation promotees hair regeneration - Google Patents
Application of the mescenchymal stem cell excretion body in the pharmaceutical preparation that preparation promotees hair regeneration Download PDFInfo
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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
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
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Abstract
The invention discloses a kind of application of mescenchymal stem cell excretion body in the pharmaceutical preparation that preparation promotees hair regeneration.The mescenchymal stem cell is prepared by following methods: the separation and Extraction mescenchymal stem cell from umbilical cord, is then obtained by originally culture and secondary culture.Application of the mescenchymal stem cell excretion body provided by the invention in the pharmaceutical preparation that preparation promotees hair regeneration, using mescenchymal stem cell excretion body, skin texture at alopecia areata can be improved, promote the hair follicle at alopecia areata in skin histology in resting stage to growth period conversion, promotes hair regeneration at alopecia areata.Mescenchymal stem cell excretion body will have broad application prospects in the pharmaceutical preparation that preparation promotees hair regeneration.
Description
Technical field
The invention belongs to the purposes technical fields of mescenchymal stem cell excretion body, specifically, it is dry to be related to a kind of mesenchyma
Application of the cell excretion body in the pharmaceutical preparation that preparation promotees hair regeneration.
Background technique
There are many diseases for leading to trichomadesis, and one of the most common is alopecia areata.Alopecia areata (Alopecia areata) is commonly called as
" alopecia areata " is a kind of patchy alopecia of limitation occurred suddenly, and local skin is normal, no conscious sympton.The cause of disease at present
Unknown, the visible hair follicle of pathologic finding is reduced, and has lymphocytic infiltration in perifollicolar, and this disease merges other autoimmunities sometimes
Property disease (Alopecia Areata:Review of Epidemiology, Clinical Features, Pathogenesis,
and New TreatmentOptions.Darwin E,Hirt PA,Fertig R,Doliner B,Delcanto G,
Jimenez JJ.Int J Trichology.2018；10(2):51-60).Most of common alopecia areatas have spontaneous recovery tendency, few
Number case repeated, so treatment is difficult, but have many therapies can be with combination therapy alopecia.Any age can be betided, but
With between twenty and fifty common, both sexes disease incidence no significant difference.Skin lesion shows as the round or non-alopecia cicatrisata of oval, on alopecia areata side
Edge Chang Kejian " exclamation mark " sample hair.Hair all or almost all falls off, referred to as whole alopecia.All hairs of whole body (including body
Hair) all fall off, it is referred to as general de-, it is also shown row alopecia of crawling.Other exceptions are not present in addition to no hair in lesion skin.Have at present
Document report can make alopecia areata model by the method for partial smearing imiquimod in C3H/HeJ mouse, at present for alopecia areata
It there is no effective treatment method.
Mescenchymal stem cell (Mesenchymal Stem Cells, MSC) be it is a kind of from a wealth of sources, have height self more
The multipotential stem cell of new ability and multi-lineage potential, experimental study, which has proven to MSC, to be divided into are as follows: bone, cartilage, fat etc. are more
Kind of cell, has been applied to a variety of diseases of clinical treatment at present, the disease for the treatment of include degenerated joint lesion (such as: bone joint
Inflammation, degenerative disc disease)；Wound (such as: cross ligament damage, rotator cuff tear etc.) (Barry F, Murphy
M.Mesenchymal stem cells in joint disease and repair.Nat Rev Rheumatol.2013,9
(10): 584-94.), however its therapy mechanism is still not clear.MSC adhesion is strong, can be adherent rapidly, there is the ability of fast breeding.
Be in shuttle shape or triangle after cell is adherent, similar fibroblast, repeatedly after passage cellular morphology without significantly changing.MSC's
Screening and identification be usually a group-specific antigen label be positive, comprising: SH2, SH3, CD29, CD44, CD90, CD106,
CD33.Hematopoietic lineage antigen is negative, including CD34, CD45, CD14.In addition, MSC also has immunogenicity low and immunosupress
Effect.But directly adopt MSC carry out clinical treatment still have biggish immunological rejection risk (Wang Y, Chen X,
Cao W,Shi Y.Plasticity of mesenchymal stem cells in immunomodulation:
pathological and therapeutic implications.Nat Immunol.201415(11):1009-16)。
Excretion body (exosomes) is the vesicles that the diameter that a variety of living cells are secreted in a manner of sprouting is about 20-140nm
Body can secrete very much (the Marb á n E.The Secret Life of Exosomes:What Bees of cell category of excretion body
Can Teach Us About Next-Generation Therapeutics.J Am Coll Cardiol.2018Jan 16；
71 (2): 193-200), not only there is its specific proteins molecule from the excretion body of different cells, but also include its row
The key molecule for making function has in many physiology such as the generation of wound repair, inflammatory reaction and tumour and transfer and pathologic process
Important function.There is no literature reported on the applications in relation to excretion body in hair regeneration at present.
Summary of the invention
The object of the present invention is to provide a kind of mescenchymal stem cell excretion bodies in the pharmaceutical preparation that preparation promotees hair regeneration
Application.
To achieve the goals above, The technical solution adopted by the invention is as follows:
The first aspect of the invention provides the drug system that a kind of mescenchymal stem cell excretion body promotees hair regeneration in preparation
Application in agent.
The mescenchymal stem cell excretion body is obtained by P3-P5 for mescenchymal stem cell culture.
The mescenchymal stem cell excretion body is prepared by following methods: the mesenchyma for being passaged to 3-5 generation is dry thin
Born of the same parents plant in culture dish, when cell fusion is up to 60~80%, cleans cell, culture of the replacement without excretion body serum with PBS
Base continues after cultivating 48-72h, collects cell conditioned medium, centrifugation removal cell or cell fragment, extracts to obtain with supercentrifugation
Mescenchymal stem cell excretion body.
The mescenchymal stem cell can be selected from fresh umbilical cord culture people's umbilical cord mesenchymal stem cells, and detailed step is referring to embodiment
1, it can also be bought by ScienCell company, the U.S..
The source for mesenchymal stem cells is in umbilical cord.
The mescenchymal stem cell is prepared by following methods:
Then the separation and Extraction mescenchymal stem cell from umbilical cord is obtained by originally culture and secondary culture.
The mescenchymal stem cell is prepared by following methods:
Aseptically, umbilical cord is cut into segment, sufficiently washed, removed arteriovenous, umbilical cord segment is shredded, training is placed in
It supports in ware, the low sugar DMEM culture solution dual anti-containing fetal calf serum and green chain is added, is placed in CO2Culture, is then added in incubator
Culture solution is replaced culture solution after 3d, is changed the liquid once every 3d, reaches 70-80% to mescenchymal stem cell fusion, is digested with pancreatin,
Carry out secondary culture.
A kind of pharmaceutical preparation for treating alopecia areata, the pharmaceutical preparation include above-mentioned mescenchymal stem cell excretion body.
The pharmaceutical preparation further includes medicinal diluent, and those skilled in the art can be known by this field professional book
These pharmaceutically acceptable pharmaceutical carrier, auxiliary material or diluents.
The concentration of mescenchymal stem cell excretion body is 50ug/ml in the pharmaceutical preparation.
For the pharmaceutical preparation by administration in subcutaneous injection, dosage is 50ug skin every square centimeter.
The present invention, which is experimentally confirmed umbilical cord mesenchymal stem cells excretion body (UMSC-exo), can enter target organ: thin with living
Born of the same parents' coloring agent CM-Dil marks UMSC-exo respectively, and skin at alopecia areata is injected into after collection, after 72 hours, frozen section materials dyeing
The case where entering target organ with fluorescence microscope UMSC-exo, as a result, it has been found that the UMSC-exo of red fluorescence label can succeed
Enter skin histology at alopecia areata.
The present invention can be obviously promoted the hair that resting stage is chronically at alopecia areata by a series of experiments proof UMSC-exo
Capsule promotes hair regeneration to growth period follicle switches.UMSC-exo promotes the table of certain cell factors in skin histology at alopecia areata
It reaches, promotes hair follicle growth, promote the resting stage hair follicle restoration ecosystem of exclamation mark sample being located in dermal layer of the skin and enter growth
Phase, hair follicle gradually increase, overstriking, and hair follicle root enters subcutaneous tissue, hair regeneration.Therefore, UMSC-exo treats spot in preparation
To have broad application prospects in bald pharmaceutical preparation.
Due to the adoption of the above technical scheme, the present invention has the following advantages and beneficial effects:
Application of the mescenchymal stem cell excretion body provided by the invention in the pharmaceutical preparation that preparation promotees hair regeneration, preparation
The raw material sources of UMSC-exo are umbilical cords, and preparation cost is cheap, and acquisition is convenient, are non-invasive operation, umbilical cord abundance can
To carry out large-scale production；The method of separation and Extraction UMSC-exo is simple, effective.
Application of the mescenchymal stem cell excretion body provided by the invention in the pharmaceutical preparation that preparation promotees hair regeneration, uses
Mescenchymal stem cell excretion body can improve skin texture at alopecia areata, promote to be in resting stage at alopecia areata in skin histology
Hair follicle promotes hair regeneration at alopecia areata to growth period conversion.Mescenchymal stem cell excretion body promotees the drug of hair regeneration in preparation
To have broad application prospects in preparation.
Detailed description of the invention
Fig. 1 is the light microscopic figure of umbilical cord mesenchymal stem cells (UMSC) in embodiment 1.
Fig. 2 is the Western blot qualification result of umbilical cord mesenchymal stem cells excretion body (UMSC-exo) in embodiment 1
Figure.
Fig. 3 is the qualification result figure of the size of umbilical cord mesenchymal stem cells excretion body (UMSC-exo) diameter in embodiment 1.
Fig. 4 is the miniflow formula scatter plot of umbilical cord mesenchymal stem cells excretion body (UMSC-exo) qualification result in embodiment 1.
Fig. 5 is that injection 2 umbilical cord mesenchymal stem cells excretion body (UMSC-exo) of embodiment promotes hair regeneration situation afterwards
Experimental result picture.
Fig. 6 is that 2 umbilical cord mesenchymal stem cells excretion body (UMSC-exo) of embodiment enters at alopecia areata in skin tissue cell
Experimental result picture.
Fig. 7 be embodiment 2 inject umbilical cord mesenchymal stem cells excretion body (UMSC-exo) afterwards EGF in skin at alopecia areata,
The experimental result picture of the expression of versican.
Fig. 8 is that embodiment 2 injects umbilical cord mesenchymal stem cells excretion body (UMSC-exo) skin HE dyeing at alopecia areata afterwards
Experimental result picture.
Specific embodiment
In order to illustrate more clearly of the present invention, below with reference to preferred embodiment, the present invention is described further.Ability
Field technique personnel should be appreciated that following specifically described content is illustrative and be not restrictive, this should not be limited with this
The protection scope of invention.
Instrument and reagent used in the present invention, material are well known to those skilled in the art, and can pass through commercial undertaking
Purchase obtains.Method used in the present invention, such as HE dyeing, Western blot are method well known in the art, can be led to
The description for crossing textbook or pertinent literature carries out, and repeats no more, is described in description of the invention, describes in the reference present invention
Method carry out.
Embodiment 1
The separation of mescenchymal stem cell excretion body is identified
1. mescenchymal stem cell (MSC) being separately cultured and identifying
Umbilical cord comes from Changhai obstetrics and gynecology hospital, and originally culture obtains mescenchymal stem cell.It is mounted in after acquisition umbilical cord and is contained
Have sterile phosphate buffer PBS (Gibco, article No.: C14190500BT) or low sugar DMEM culture medium (Gibco, article No.:
In 50ml centrifuge tube 11885-084), operate as early as possible.After PBS repeated flushing, it is soaked in containing mycillin (Gibco, goods
Number: low sugar-DMEM (mycillin concentration is 100IU/ml) 15140-122) about 5 minutes is cut into diameter about 1.5mm size
Tissue block；Containing 10% fetal calf serum low sugar-DMEM nutrient solution, 5%CO2, 37 DEG C of saturated humidity cultures；It is visible after 3-5 days
Mescenchymal stem cell climbs out of.
The mescenchymal stem cell is prepared by following methods:
Aseptically, the umbilical cord for taking full-term pregnancy natural labor healthy fetus, cuts about 3cm long, and merging contains 100IU/
In the low sugar DMEM culture solution of ml mycillin, culture is initially separated in 2 hours as far as possible.It is sufficiently rinsed with PBS and removes navel
Tissue, is then cut into the bulk of diameter about 1.5mm size by band arteriovenous.Fritter is spread evenly across culture dish (BD
Falcon, article No.: 353002) bottom is added and contains 10% fetal calf serum base (Gibco, article No.: 10099-141) and 100IU/ml
The dual anti-low sugar DMEM culture solution of green chain, gently overturns culture dish, culture dish bottom is made gently to be placed on CO upward2In incubator
(Thermo, ORMA 3111).After 2-4 hours, it is attached at ware bottom to fritter, culture dish is slowly turningly laid flat, is added a small amount of
Culture medium, stationary culture, movement is light in the process, forbids to shake.Culture medium should not be more when starting to cultivate, to keep tissue block wet
Profit, the fluid infusion again after culture 24 hours.It to handle with care when mobile and observation when being observed in 3 days, not go to move as far as possible
Culture dish, in favor of the adherent of tissue block and growth.On the one hand interchangeable liquid at culture 3-5 days removes metabolite and floats small
On the one hand toxic effect caused by block supplements nutrition needed for cell is grown.After mescenchymal stem cell fusion reaches 70-80%,
It is digested with pancreatin (Gibco, article No.: 25200-056), carries out secondary culture.
Cell of the passage in 3 generations is taken to carry out subsequent operation, umbilical cord mesenchymal stem cells are identified as shown in Figure 1, by P4 for MSC
It is placed in optical microphotograph under the microscope, cellular morphology is in shuttle shape or short triangle, and after 100% fusion, circinate growth is presented in cell.
The result is shown in Figure 1, Fig. 1 are the light microscopic figures of umbilical cord mesenchymal stem cells (UMSC) in embodiment 1.It is filled between 4th generation people's bleeding of the umbilicus of culture
Matter stem cell expresses CD166, CD44, CD29, CD90, and CD45, CD34 are negative.Fig. 1 result explanation obtained purity compared with
High UMSC.
2. the separation of mescenchymal stem cell excretion body
The MSC kind in P3 generation is planted in culture dish, when cell fusion is up to 60%, clean cell with PBS, replacement is without excretion
The culture medium of body serum, continues after cultivating 48-72h, collects cell conditioned medium, is centrifuged 10min with the revolving speed of 2000rpm, then with
The revolving speed of 10000rpm is centrifuged 30min, removes cell or cell fragment, uses supercentrifugation separation and Extraction MSC excretion body.
3. the identification of mescenchymal stem cell excretion body (MSC-exo)
(1) Western-blot detects the expression of the specific proteins of Exosome
With kit, (Thermo, total EXO RNA/PROT ISO KIT, article No.: 4478545) are extracted Exosome
Albumen, the loading after BCA method (Abbkine, article No.: KTD3001) detection protein content, electrophoresis, transferring film.It is immersed in 5%BSA envelope
Room temperature in liquid is closed slowly to sway two hours.It is incubated for 4 DEG C of primary antibody overnight, suitable secondary antibody is selected according to primary antibody source, by corresponding ratio
Example dilution (1:1000~1:10000), room temperature jog two hours.After TBS washing, ECL luminescence reagent (Abbkine, goods are used
Number: it K22020) develops the color, as a result sees that Fig. 2, Fig. 2 are umbilical cord mesenchymal stem cells excretion bodies (UMSC-exo) in embodiment 1
Western blot qualification result figure, it can be seen from the figure that excretion body Specific marker CD81, CD63 is high-visible.
(2) diameter of nanoparticle analyzer analysis MSC excretion body
It is analyzed using nano biological grain analyser (New Zealand iZON science), after taking 500 microlitres of dilutions
Sample to be tested, and operated in advance with 0.22 zut filter by instrument specification, as a result see that Fig. 3 and Fig. 4, Fig. 3 are real
The qualification result figure of the size of umbilical cord mesenchymal stem cells excretion body (UMSC-exo) diameter in example 1 is applied, it can be seen from the figure that
The diameter mean size of UMSC-exo is 119nm.Fig. 4 is umbilical cord mesenchymal stem cells excretion body (UMSC-exo) in embodiment 1
The miniflow formula scatter plot of qualification result shows that the diameter of UMSC-exo is distributed in 80-150nm more, illustrates to have obtained and fill between umbilical cord
Matter stem cell excretion body.
Embodiment 2
The excretion body of umbilical cord mesenchymal stem cells secretion can improve skin histology microenvironment at alopecia areata, promote hair again
Raw, the requirement of NIH experimental animal is abided by all zooperies.
Experimental animal is 6 week old C3H/HeJ mouses, and male and female are unlimited, weighs about 30g, by The 2nd Army Medical College experimental animal
Center provides.
Alopecia areata model is formed using imiquimod induction C3H/HeJ mouse first, specific method refers to CN105012969A,
It is a kind of to protect and promote the drug of hair regeneration and its application at alopecia areata.
The successful mouse of alopecia areata modeling is randomly divided into 2 groups, respectively experimental group, that is, UMSC-exo injection group and control group
PBS injection group.Every group 5,1% Nembutal sodium solution is injected, is injected at alopecia areata after anesthesia with microsyringe, often
The UMSC-exo of square centimeter alopecia areata injection of skin 50ug, it is daily to observe, 3 after transplanting, draw materials within 5,7,9,14 days.
1. hair regeneration situation is observed
After UMSC-exo injection, to determine hair regeneration effect, digital camera tracks skin and hair regeneration at alopecia areata of taking pictures
Situation.As shown in figure 5, Fig. 5 is that injection 2 umbilical cord mesenchymal stem cells excretion body (UMSC-exo) of embodiment promotes hair regeneration afterwards
The experimental result picture of situation, before being above classified as injection, under be classified as injection after two weeks as a result, the results show that UMSC-exo injection group
(right side) hair regeneration situation is significantly higher than PBS injection group (left side), illustrates that UMSC-exo can significantly promote at alopecia areata hair again
It is raw.
2.UMSC-exo enters at alopecia areata in skin histology the case where cell
It draws materials after UMSC-exo injection, is drawn materials centered on injection point, diameter 1cm or so.It is small that 4% paraformaldehyde fixes 24
When, saccharose gradient dehydration, OCT embedding, freezing microtome serial section, slice thickness is 5 μm.DAPI (4', 6- are carried out to slice
Diamidino -2-phenylindone；4', 6-diamidino-2-phenylindole) dyeing after in fluorescence microscopy under the microscope, as a result
See that Fig. 6, Fig. 6 are that 2 umbilical cord mesenchymal stem cells excretion body (UMSC-exo) of embodiment enters at alopecia areata in skin tissue cell
Experimental result picture, it can be seen from the figure that UMSC-exo can be entered in the cell of hair follicle；Left figure is DAPI dyeing knot in Fig. 6
Fruit shows the nucleus in skin histology herein.The excretion body that right figure display fluorescent dye carries fluorescent dye comes into
Skin histology at alopecia areata, illustrate subcutaneous injection excretion body can enter skin histology in cell, by its carry substance,
Promote hair regeneration.
At 3.UMSC-exo promotion alopecia areata in skin histology the case where cytokine-expressing
It draws materials after UMSC-exo injection, is drawn materials centered on injection point, taken skin is carried out albumen by diameter 1cm or so
Extracting, BCA standard measure.The table for promoting the factor such as EGF and versican of hair growth is detected through Western-blot method after
Up to situation, as a result as shown in Figure 7, Fig. 7 is that embodiment 2 injects umbilical cord mesenchymal stem cells excretion body (UMSC-exo) afterwards at alopecia areata
The experimental result picture of the expression of EGF, versican in skin, it can be seen from the figure that after UMSC-exo injection, at alopecia areata
The factor expression amount for promoting hair regeneration in skin increases.
The microstructure of skin at 4.HE dyeing observation alopecia areata
It draws materials after UMSC-exo injection, takes each group alopecia areata central diameter 1cm expanses of skin, it is small that 4% paraformaldehyde fixes 24
When, paraffin embedding, serial section, slice thickness is 5 μm.Skin biopsy row HE at alopecia areata is dyed, as shown in figure 8, Fig. 8 is real
Apply example 2 inject umbilical cord mesenchymal stem cells excretion body (UMSC-exo) afterwards at alopecia areata skin HE dyeing experimental result picture, it is micro-
Find under the microscope: UMSC-exo injection group hair follicle increases, and hair follicle structure is complete, into growth period, grows downwards；Explanation
After UMSC-exo injection, skin texture is improved, and the hair regeneration of skin at alopecia areata is promoted.
The basic principles, main features and advantages of the present invention have been shown and described above.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this
The principle of invention, various changes and improvements may be made to the invention without departing from the spirit and scope of the present invention, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and its
Equivalent defines.
Claims (10)
1. a kind of application of mescenchymal stem cell excretion body in the pharmaceutical preparation that preparation promotees hair regeneration.
2. mescenchymal stem cell excretion body according to claim 1 answering in the pharmaceutical preparation that preparation promotees hair regeneration
With, it is characterised in that: the mescenchymal stem cell excretion body is obtained by P3-P5 for mescenchymal stem cell culture.
3. mescenchymal stem cell excretion body according to claim 2 answering in the pharmaceutical preparation that preparation promotees hair regeneration
With, it is characterised in that: the mescenchymal stem cell excretion body is prepared by following methods: will be passaged between 3-5 generation and is filled
Matter stem cell is planted in culture dish, when cell fusion is up to 60~80%, clean cell with PBS, replacement is without excretion body serum
Culture medium, continue after cultivating 48-72h, collect cell conditioned medium, centrifugation removal cell or cell fragment is mentioned with supercentrifugation
Obtain mescenchymal stem cell excretion body.
4. mescenchymal stem cell excretion body according to claim 3 answering in the pharmaceutical preparation that preparation promotees hair regeneration
With, it is characterised in that: the source for mesenchymal stem cells is in umbilical cord.
5. mescenchymal stem cell excretion body according to claim 4 answering in the pharmaceutical preparation that preparation promotees hair regeneration
With, it is characterised in that: the mescenchymal stem cell is prepared by following methods:
Then the separation and Extraction mescenchymal stem cell from umbilical cord is obtained by originally culture and secondary culture.
6. mescenchymal stem cell excretion body according to claim 5 answering in the pharmaceutical preparation that preparation promotees hair regeneration
With, it is characterised in that: the mescenchymal stem cell is prepared by following methods:
Aseptically, umbilical cord is cut into segment, sufficiently washed, removed arteriovenous, umbilical cord segment is shredded, culture dish is placed in
In, the low sugar DMEM culture solution dual anti-containing fetal calf serum and green chain is added, is placed in CO2Culture in incubator, then addition culture
Liquid is replaced culture solution after 3d, is changed the liquid once every 3d, reaches 70-80% to mescenchymal stem cell fusion, is digested, carried out with pancreatin
Secondary culture.
7. a kind of pharmaceutical preparation for treating alopecia areata, it is characterised in that: the pharmaceutical preparation includes any one of claim 1 to 6 institute
The mescenchymal stem cell excretion body stated.
8. the pharmaceutical preparation for the treatment of alopecia areata according to claim 7, it is characterised in that: the pharmaceutical preparation further includes pharmacy
Upper acceptable pharmaceutical carrier, auxiliary material or diluent.
9. the pharmaceutical preparation for the treatment of alopecia areata according to claim 7, it is characterised in that: mesenchyma is dry in the pharmaceutical preparation
The concentration of cell excretion body is 50ug/ml.
10. the pharmaceutical preparation for the treatment of alopecia areata according to claim 7, it is characterised in that: the pharmaceutical preparation passes through subcutaneous
Administration in injection, dosage are 50ug skin every square centimeter.
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| CN113897387A (en) * | 2021-10-09 | 2022-01-07 | 深圳市汉科生命工程有限公司 | Gene recombinant MSC with hair regeneration promoting function and preparation method and application of exosome-like nano material thereof |
| CN114159550A (en) * | 2021-11-16 | 2022-03-11 | 南京天纵易康生物科技股份有限公司 | Preparation method of microcapsule hair-growing product based on exosome growth factor |
| CN114231483A (en) * | 2021-12-21 | 2022-03-25 | 广州市新甡命科技有限公司 | Stem cell exosome and application thereof in promoting hair growth |
| CN115572709A (en) * | 2022-10-18 | 2023-01-06 | 中晶生物技术股份有限公司 | Application of three-dimensional culture mesenchymal stem cell microspheres in preparation of alopecia repairing and regenerating medicine or preparation |
| CN115814045A (en) * | 2022-11-03 | 2023-03-21 | 山东新创生物科技有限公司 | Exosome composite preparation for improving alopecia and preparation method and application thereof |
| CN113813289B (en) * | 2021-09-07 | 2023-10-27 | 深圳市莱利赛生物科技有限公司 | Preparation method of hair follicle generation promoting liquid based on umbilical cord mesenchymal stem cell exosomes |
| CN117100673A (en) * | 2023-10-23 | 2023-11-24 | 广州优特佳生物科技发展有限公司 | Animal umbilical cord extract for regulating hair follicle cycle, and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130209528A1 (en) * | 2010-10-18 | 2013-08-15 | Agency For Science, Technology And Research | Use of exosomes to promote or enhance hair growth |
| WO2015048844A1 (en) * | 2013-10-02 | 2015-04-09 | Xosoma Pty Ltd | A method of producing exosomes |
| CN106511385A (en) * | 2016-12-09 | 2017-03-22 | 济南万泉生物技术有限公司 | Medicine for treating hair loss caused by chemotherapy |
-
2018
- 2018-11-21 CN CN201811392587.XA patent/CN109432127A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130209528A1 (en) * | 2010-10-18 | 2013-08-15 | Agency For Science, Technology And Research | Use of exosomes to promote or enhance hair growth |
| WO2015048844A1 (en) * | 2013-10-02 | 2015-04-09 | Xosoma Pty Ltd | A method of producing exosomes |
| CN106511385A (en) * | 2016-12-09 | 2017-03-22 | 济南万泉生物技术有限公司 | Medicine for treating hair loss caused by chemotherapy |
Non-Patent Citations (3)
| Title |
|---|
| AMIT N. PATEL等: "《Mesenchymal stem cell population isolated from the subepithelial layer of umbilical cord tissue》", 《CELL TRANSPLANTATION》 * |
| TINGFEN LI等: "《Exosomes derived from human umbilical cord mesenchymal stem cells alleviate liver fibrosis》", 《STEM CELLS AND DEVELOPMENT》 * |
| 杨向荣等: "《人脐带间充质干细胞来源外泌体的生物学特性研究》", 《华中科技大学学报（医学版）》 * |
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| CN114159550A (en) * | 2021-11-16 | 2022-03-11 | 南京天纵易康生物科技股份有限公司 | Preparation method of microcapsule hair-growing product based on exosome growth factor |
| CN114231483A (en) * | 2021-12-21 | 2022-03-25 | 广州市新甡命科技有限公司 | Stem cell exosome and application thereof in promoting hair growth |
| CN115572709A (en) * | 2022-10-18 | 2023-01-06 | 中晶生物技术股份有限公司 | Application of three-dimensional culture mesenchymal stem cell microspheres in preparation of alopecia repairing and regenerating medicine or preparation |
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| CN117100673A (en) * | 2023-10-23 | 2023-11-24 | 广州优特佳生物科技发展有限公司 | Animal umbilical cord extract for regulating hair follicle cycle, and preparation method and application thereof |
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