CN113750118A - Application of human umbilical vein endothelial cells in preparation of anti-aging product, anti-aging freeze-dried powder and anti-aging solution - Google Patents
Application of human umbilical vein endothelial cells in preparation of anti-aging product, anti-aging freeze-dried powder and anti-aging solution Download PDFInfo
- Publication number
- CN113750118A CN113750118A CN202110911754.2A CN202110911754A CN113750118A CN 113750118 A CN113750118 A CN 113750118A CN 202110911754 A CN202110911754 A CN 202110911754A CN 113750118 A CN113750118 A CN 113750118A
- Authority
- CN
- China
- Prior art keywords
- aging
- umbilical vein
- human umbilical
- vein endothelial
- skin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 210000002889 endothelial cell Anatomy 0.000 title claims abstract description 104
- 210000003606 umbilical vein Anatomy 0.000 title claims abstract description 97
- 230000003712 anti-aging effect Effects 0.000 title claims abstract description 53
- 239000000843 powder Substances 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 210000004204 blood vessel Anatomy 0.000 claims abstract description 18
- 238000007920 subcutaneous administration Methods 0.000 claims abstract description 13
- 239000002932 luster Substances 0.000 claims abstract description 9
- 230000009759 skin aging Effects 0.000 claims description 19
- WQZGKKKJIJFFOK-SVZMEOIVSA-N (+)-Galactose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-SVZMEOIVSA-N 0.000 claims description 16
- 239000008176 lyophilized powder Substances 0.000 claims description 14
- 108010038807 Oligopeptides Proteins 0.000 claims description 13
- 102000015636 Oligopeptides Human genes 0.000 claims description 13
- 239000000243 solution Substances 0.000 claims description 13
- 238000007710 freezing Methods 0.000 claims description 11
- 230000008014 freezing Effects 0.000 claims description 11
- 239000011550 stock solution Substances 0.000 claims description 10
- 238000010257 thawing Methods 0.000 claims description 9
- 239000002904 solvent Substances 0.000 claims description 5
- 238000004108 freeze drying Methods 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims 1
- 230000032683 aging Effects 0.000 abstract description 13
- 230000008859 change Effects 0.000 abstract description 9
- 210000003491 skin Anatomy 0.000 description 91
- 210000004027 cell Anatomy 0.000 description 53
- 239000000047 product Substances 0.000 description 35
- 241000699666 Mus <mouse, genus> Species 0.000 description 16
- 239000002504 physiological saline solution Substances 0.000 description 16
- 210000003954 umbilical cord Anatomy 0.000 description 16
- 238000011282 treatment Methods 0.000 description 13
- 239000006285 cell suspension Substances 0.000 description 10
- 210000000130 stem cell Anatomy 0.000 description 10
- 241000699670 Mus sp. Species 0.000 description 9
- 238000005406 washing Methods 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- 238000001514 detection method Methods 0.000 description 6
- 238000010438 heat treatment Methods 0.000 description 6
- 229960001134 von willebrand factor Drugs 0.000 description 6
- OZFAFGSSMRRTDW-UHFFFAOYSA-N (2,4-dichlorophenyl) benzenesulfonate Chemical compound ClC1=CC(Cl)=CC=C1OS(=O)(=O)C1=CC=CC=C1 OZFAFGSSMRRTDW-UHFFFAOYSA-N 0.000 description 5
- 239000012591 Dulbecco’s Phosphate Buffered Saline Substances 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 102100024616 Platelet endothelial cell adhesion molecule Human genes 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 108010047303 von Willebrand Factor Proteins 0.000 description 5
- 102100036537 von Willebrand factor Human genes 0.000 description 5
- 230000007910 cell fusion Effects 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 230000037394 skin elasticity Effects 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 238000009777 vacuum freeze-drying Methods 0.000 description 4
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 230000003679 aging effect Effects 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 210000004207 dermis Anatomy 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 230000032258 transport Effects 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 2
- TXFPEBPIARQUIG-UHFFFAOYSA-N 4'-hydroxyacetophenone Chemical compound CC(=O)C1=CC=C(O)C=C1 TXFPEBPIARQUIG-UHFFFAOYSA-N 0.000 description 2
- 108010072135 Cell Adhesion Molecule-1 Proteins 0.000 description 2
- 102100024649 Cell adhesion molecule 1 Human genes 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- POJWUDADGALRAB-UHFFFAOYSA-N allantoin Chemical compound NC(=O)NC1NC(=O)NC1=O POJWUDADGALRAB-UHFFFAOYSA-N 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000000249 desinfective effect Effects 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000007599 discharging Methods 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 210000001339 epidermal cell Anatomy 0.000 description 2
- 230000001815 facial effect Effects 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 230000007480 spreading Effects 0.000 description 2
- 238000003892 spreading Methods 0.000 description 2
- 230000002792 vascular Effects 0.000 description 2
- 230000035899 viability Effects 0.000 description 2
- 229940015975 1,2-hexanediol Drugs 0.000 description 1
- POJWUDADGALRAB-PVQJCKRUSA-N Allantoin Natural products NC(=O)N[C@@H]1NC(=O)NC1=O POJWUDADGALRAB-PVQJCKRUSA-N 0.000 description 1
- KWIUHFFTVRNATP-UHFFFAOYSA-N Betaine Natural products C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 description 1
- 102000029816 Collagenase Human genes 0.000 description 1
- 108060005980 Collagenase Proteins 0.000 description 1
- 206010013786 Dry skin Diseases 0.000 description 1
- 102000016942 Elastin Human genes 0.000 description 1
- 108010014258 Elastin Proteins 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- BIVBRWYINDPWKA-VLQRKCJKSA-L Glycyrrhizinate dipotassium Chemical compound [K+].[K+].O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@H]1CC[C@]2(C)[C@H]3C(=O)C=C4[C@@H]5C[C@](C)(CC[C@@]5(CC[C@@]4(C)[C@]3(C)CC[C@H]2C1(C)C)C)C(O)=O)C([O-])=O)[C@@H]1O[C@H](C([O-])=O)[C@@H](O)[C@H](O)[C@H]1O BIVBRWYINDPWKA-VLQRKCJKSA-L 0.000 description 1
- 241000208680 Hamamelis mollis Species 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- PWKSKIMOESPYIA-BYPYZUCNSA-N L-N-acetyl-Cysteine Chemical compound CC(=O)N[C@@H](CS)C(O)=O PWKSKIMOESPYIA-BYPYZUCNSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- KWIUHFFTVRNATP-UHFFFAOYSA-O N,N,N-trimethylglycinium Chemical compound C[N+](C)(C)CC(O)=O KWIUHFFTVRNATP-UHFFFAOYSA-O 0.000 description 1
- CAHKINHBCWCHCF-UHFFFAOYSA-N N-acetyltyrosine Chemical compound CC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 CAHKINHBCWCHCF-UHFFFAOYSA-N 0.000 description 1
- 101710160107 Outer membrane protein A Proteins 0.000 description 1
- 108010069381 Platelet Endothelial Cell Adhesion Molecule-1 Proteins 0.000 description 1
- 241000207929 Scutellaria Species 0.000 description 1
- 229920002385 Sodium hyaluronate Polymers 0.000 description 1
- 101710127774 Stress response protein Proteins 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 229960004308 acetylcysteine Drugs 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 229960000458 allantoin Drugs 0.000 description 1
- 229940069521 aloe extract Drugs 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229960003237 betaine Drugs 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 229960002424 collagenase Drugs 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 229940101029 dipotassium glycyrrhizinate Drugs 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 230000037336 dry skin Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 229920002549 elastin Polymers 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 229940084760 glycyrrhiza inflata root extract Drugs 0.000 description 1
- FHKSXSQHXQEMOK-UHFFFAOYSA-N hexane-1,2-diol Chemical compound CCCCC(O)CO FHKSXSQHXQEMOK-UHFFFAOYSA-N 0.000 description 1
- 238000003125 immunofluorescent labeling Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 235000021590 normal diet Nutrition 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000008823 permeabilization Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 230000019612 pigmentation Effects 0.000 description 1
- 230000004983 pleiotropic effect Effects 0.000 description 1
- 239000003761 preservation solution Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000007665 sagging Methods 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 229940010747 sodium hyaluronate Drugs 0.000 description 1
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 description 1
- 229940084750 sophora flavescens root extract Drugs 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- 230000008728 vascular permeability Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 229940118846 witch hazel Drugs 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
- A61K35/51—Umbilical cord; Umbilical cord blood; Umbilical stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- 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
- A61P17/16—Emollients or protectives, e.g. against radiation
-
- 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
- A61P17/18—Antioxidants, e.g. antiradicals
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Cell Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Developmental Biology & Embryology (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Dermatology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Toxicology (AREA)
- Inorganic Chemistry (AREA)
- Hematology (AREA)
- Reproductive Health (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Virology (AREA)
- Zoology (AREA)
- Cosmetics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention relates to application of human umbilical vein endothelial cells in preparation of anti-aging products, anti-aging freeze-dried powder and anti-aging solution, and belongs to the technical field of anti-aging products. The human umbilical vein endothelial cells can be used for preparing an anti-aging product, and can improve the luster of aging skin, increase the thickness of the skin, reduce the number of subcutaneous blood vessels, increase the water content of the skin and/or reduce the oil content of the skin, and change the skin from rough to soft.
Description
Technical Field
The invention relates to the technical field of anti-aging products, in particular to application of human umbilical vein endothelial cells in preparation of anti-aging products, anti-aging freeze-dried powder and anti-aging solution.
Background
Skin aging is a phenomenon of skin aging caused by various factors. Skin aging is mainly manifested by dry skin, sagging skin, appearance of wrinkles, hyperkeratosis, pigmentation, reduction in elasticity and softness, and the like. The main inducements are the natural decline of the functions of the skin accessory organs, the slow metabolism of the facial skin, and the adverse life factors (staying up, smoking and drinking) of modern people not only accelerate but also directly cause the aging of the skin. With the development of society and the improvement of living standard of people, the anti-aging treatment of skin becomes a hot point of medical cosmetology.
The conventional anti-aging skin care product mainly comprises chemical components, and is only suitable for basic water replenishing and water locking of young skin, so that the problem of skin aging cannot be improved, and the requirement of aging skin in young state is met.
Stem cells are a unique cell population with self-renewal and differentiation properties, and are widely used in clinical anti-aging studies due to their pleiotropic, easily separable, and multilineage differentiation properties. In recent years, research on the clinical use of stem cells for resisting skin aging has increased. At present, research on stem cell anti-aging is mostly focused on mesenchymal stem cells and freeze-dried powder or secreted factors thereof. The living cells injected into veins and faces are mainly used, and the fresh cells are used, so the transportation requirement is high, and the transportation cost is high; secondly, the activity of the fresh cells is difficult to completely preserve, the prepared fresh cells generally need to be used immediately within 4h, and the activity and the performance are reduced the longer the time is; through the research of using the umbilical cord mesenchymal stem cell freeze-dried powder to resist skin aging, the crowd sample is younger and mostly concentrated between 30 and 50 years old; in addition, the secretion factor for resisting skin aging can overcome the defects of inconvenient use, increased transportation and storage cost and the like, but cannot completely obtain the biological active factor of the cell.
In summary, there is a need to develop a new class of stem cell derived cosmetic products that are more suitable for the needs of aging skin youthfulness to provide a new means of anti-aging treatment of skin.
Disclosure of Invention
The invention aims to provide application of human umbilical vein endothelial cells in preparation of anti-aging products, anti-aging freeze-dried powder and anti-aging solution. The human umbilical vein endothelial cells can be used for preparing an anti-aging product, and can improve the luster of aging skin, increase the thickness of the skin, reduce the number of subcutaneous blood vessels, increase the water content of the skin and/or reduce the oil content of the skin, and change the roughness of the skin into the fineness; the prepared product has good anti-aging effect, is convenient to use, transport and store, is more suitable for the anti-aging of the skin of the aged, has less cell consumption, and can meet the young requirement of the aged skin.
The invention provides an application of human umbilical vein endothelial cells in preparing an anti-aging product.
The invention also provides application of the human umbilical vein endothelial cells in preparing products for resisting skin aging.
The invention also provides application of the human umbilical vein endothelial cells in preparing products for resisting skin aging caused by D-galactose.
The invention also provides application of the human umbilical vein endothelial cells in preparing a product for improving the luster of aged skin.
The invention also provides application of the human umbilical vein endothelial cells in preparing a product for increasing the thickness of aged skin.
The invention also provides application of the human umbilical vein endothelial cells in preparing a product for reducing the number of subcutaneous blood vessels of aged skin.
The invention also provides application of the human umbilical vein endothelial cells in preparing a product for increasing the water content of aged skin and/or reducing the oil content of the skin.
The invention also provides application of the human umbilical vein endothelial cells in preparing a product for improving the roughness of aged skin.
The invention also provides anti-aging human umbilical vein endothelial cell freeze-dried powder, and the preparation method of the freeze-dried powder comprises the following steps:
the invention also provides freeze-dried powder of the human umbilical vein endothelial cells, which is obtained by repeatedly freezing and thawing the human umbilical vein endothelial cells and freeze-drying the human umbilical vein endothelial cells in vacuum.
The invention also provides an anti-aging human umbilical vein endothelial cell freeze-dried powder solution, which is obtained by mixing oligopeptide stock solution serving as a solvent and freeze-dried powder serving as a solute according to the technical scheme.
The invention provides an application of human umbilical vein endothelial cells in preparing an anti-aging product. The human umbilical vein endothelial cell product has wide application prospect due to good anti-skin aging effect, and can be applied to 1) clinical application: medical cosmetic orientation; 2) an anti-aging skin care product. The human umbilical vein endothelial cells can be used for preparing an anti-aging product, and can improve the luster of aging skin, increase the thickness of the skin, reduce the number of subcutaneous blood vessels, increase the water content of the skin and/or reduce the oil content of the skin, and change the skin from rough to soft. The product prepared by the application of the invention has good anti-aging effect, is convenient to use, transport and store, is more suitable for the anti-aging of the skin of the aged, has less cell consumption, and can meet the young state requirement of the aged skin.
Drawings
FIG. 1 is a diagram of the identification and purity detection of clinical grade human umbilical vein endothelial cells provided by the present invention; wherein, A is the result of specific marker positive expression of Von Willebrand Factor (Von Willebrand Factor) of endothelial cells; b is the purity result of endothelial cells detected by flow detection of CD3 l antigen expression;
FIG. 2 is the change result of mouse body weight within 4 weeks of clinical grade human umbilical vein endothelial cell lyophilized powder intervention provided by the present invention;
FIG. 3 shows the results of the anti-aging effect of clinical grade lyophilized powder of human umbilical vein endothelial cells on mouse skin.
Detailed Description
The invention provides an application of human umbilical vein endothelial cells in preparing an anti-aging product. The human umbilical vein endothelial cells are preferably clinical-grade human umbilical vein endothelial cells, and the purity is preferably 99.9%. The method for separating and culturing the clinical-grade human umbilical vein endothelial cells is not particularly limited, and the conventional method for separating and culturing the human umbilical vein endothelial cells, which is well known to those skilled in the art, can be adopted. The human umbilical vein endothelial cells are applied to preparing the anti-aging product, and can play a role in anti-aging. The effect on the anti-aging improvement of aged skin is remarkable. The aging of the present invention preferably means a population aged about 60 years.
The invention also provides application of the human umbilical vein endothelial cells in preparing products for resisting skin aging. The human umbilical vein endothelial cells can improve the luster of aged skin, increase the thickness of the skin, reduce the number of subcutaneous blood vessels, increase the moisture content of the skin and/or reduce the oil content of the skin, change the skin from rough to soft and increase the elasticity of the skin.
The invention also provides application of the human umbilical vein endothelial cells in preparing products for resisting skin aging caused by D-galactose. The human umbilical vein endothelial cells can resist skin aging caused by D-galactose, improve skin luster, increase skin thickness, reduce subcutaneous blood vessel number, increase skin water content and/or reduce skin oil content, change skin from rough to soft, and increase skin elasticity.
The invention also provides application of the human umbilical vein endothelial cells in preparing a product for improving the luster of aged skin. The test result shows that the skin is bright and glossy after being treated by the endothelial cells of the umbilical vein of human.
The invention also provides application of the human umbilical vein endothelial cells in preparing a product for increasing the thickness of aged skin. The test result shows that the skin becomes thicker after being treated by the endothelial cells of the umbilical vein of the human.
The invention also provides application of the human umbilical vein endothelial cells in preparing a product for reducing the number of subcutaneous blood vessels of aged skin. Test results show that subcutaneous blood vessels are remarkably reduced after the treatment of human umbilical vein endothelial cells.
The invention also provides application of the human umbilical vein endothelial cells in preparing a product for increasing the water content of aged skin and/or reducing the oil content of the skin. Test results show that after being treated by human umbilical vein endothelial cells, the water content of the skin is increased, the oil content is reduced, and the skin elasticity is increased.
The invention also provides application of the human umbilical vein endothelial cells in preparing a product for improving the roughness of aged skin. Test results show that the skin state changes from rough to delicate after being treated by human umbilical vein endothelial cells.
The invention also provides anti-aging human umbilical vein endothelial cell freeze-dried powder, and the preparation method of the freeze-dried powder comprises the following steps:
and (3) repeatedly freezing and thawing the human umbilical vein endothelial cells, and freeze-drying in vacuum to obtain the human umbilical vein endothelial cell freeze-dried powder.
Specifically, the invention preferably expands human umbilical vein endothelial cells, and the harvested cells are resuspended in physiological saline and frozen into solid state at-80 ℃; thawing, diluting with water to obtain cell suspension, freezing at-80 deg.C, and repeating the above steps to obtain repeatedly frozen and thawed cell suspension; and (4) carrying out vacuum freeze-drying on the repeatedly frozen and thawed cells to obtain the human umbilical vein endothelial cell freeze-dried powder.
The invention expands human umbilical vein endothelial cells, and the harvested cells are resuspended in physiological saline and frozen into solid at-80 ℃. The invention preferably selects human umbilical vein endothelial cells from primary passage to P5 for expansion. The present invention preferably harvests 1000 million cells resuspended in 10mL of physiological saline. The conditions of-80 ℃ according to the invention are preferably provided by a refrigerator.
The invention is unfrozen, diluted by adding water to obtain cell suspension, frozen at-80 ℃, and the step is repeated to obtain the cells which are repeatedly frozen and thawed. According to the invention, 10 times of the volume of water is preferably added, and the water is preferably double distilled water. The-80 ℃ condition according to the invention is preferably provided by a-80 ℃ refrigerator. The freezing is preferably overnight freezing, and more preferably freezing for 8-12 h. The number of times of repeated freeze thawing is preferably 3-5, and more preferably 3.
After repeated freeze thawing, the invention carries out vacuum freeze drying on the cells subjected to repeated freeze thawing to obtain the human umbilical vein endothelial cell freeze-dried powder. The human umbilical vein endothelial cell freeze-dried powder is preferably stored at 4 ℃ after being prepared. Specifically, the vacuum freeze-drying preferably comprises the following steps: starting a vacuum freeze dryer, and freezing the cell suspension to-45 ℃; then starting a freeze dryer cold trap to refrigerate to-45 ℃, and vacuumizing to 20 Pa; opening the box body and heating to-20 deg.C, and maintaining for 240 min; continuously heating to 1 deg.C and maintaining for 240 min; continuously heating to 30 ℃ and preserving heat for 300 min; discharging at 36 ℃ after the detection is qualified; and (6) filling, inspecting and warehousing.
When the human umbilical vein endothelial cell freeze-dried powder is used, a solvent is preferably used for dilution, and after dilution, each milliliter of solution preferably contains 2000-50000 human umbilical vein endothelial cells.
The human umbilical vein endothelial cell freeze-dried powder has an anti-aging function and strong anti-skin aging activity, can improve the luster of aged skin, increase the thickness of the skin, reduce the number of subcutaneous blood vessels, increase the water content of the skin and/or reduce the oil content of the skin, and changes the roughness of the skin into the fineness of the skin. The purity of the human umbilical vein endothelial cells obtained by the preparation method is as high as 99.9 percent, and the human umbilical vein endothelial cells are clinical-grade human umbilical vein endothelial cells, have the advantages of easy obtainment, no ethical dispute and noninvasive material drawing, and are convenient to use, store and transport; at present, the clinical research of stem cell anti-aging in the market mainly takes intravenous and facial injection of living cells, and because fresh cells are used, the transportation requirement is high, and the transportation cost is increased; secondly, the viability of the fresh cells cannot be completely preserved, and the prepared fresh cells generally need to be used immediately within 4h, and the longer the time is, the viability and the performance are reduced. The invention combines a vacuum freeze-drying method, freezes the water contained in the stem cells at low temperature, and then dries the stem cells in a vacuum environment, so that the water is directly sublimated into water vapor from a solid state and is discharged from a cell product to dry all active factors of the cells, thereby effectively preventing the change of the physicochemical and biological properties of the cells, causing the damage to the biological tissues and the structures and the characteristics of the cells to be small, leading the cells to rapidly enter a dormant state, and effectively protecting the stability of the effective components of the cells and the biological efficacy of the active factors; secondly, the human umbilical vein endothelial cell freeze-dried powder is loose in shape and basically not changed in color after being dried, and can be quickly dissolved after being added with water and restore the physical and chemical properties and biological activity of the original aqueous solution. In addition, because the drying is carried out under the vacuum condition, the method has good protection effect on some substances which are easy to oxidize. Finally, the freeze-dried powder has very low water content after freeze-drying, so that the stability of the product is improved, the chance of pollution is reduced, the transportation is convenient, and the storage life of the product is prolonged.
At present, research on human umbilical vein endothelial cell anti-aging does not appear, and the research on stem cell anti-aging is mostly concentrated on mesenchymal stem cells and freeze-dried powder or secretion factors thereof, and although the secretion factors can overcome the defects of inconvenient use, increased transportation and storage costs and the like, the biological activity factors of the cells cannot be completely obtained; the research on the anti-skin aging by using the umbilical cord mesenchymal stem cell freeze-dried powder has the advantages that the population sample is younger, the population sample is mostly concentrated between 30 and 50 years old, and the required number of cells is large (100 ten thousand/mL). The lyophilized powder of human umbilical vein endothelial cells can enable the skin on the back of a 14-month-old subacute aging model (D-galactose induction) mouse (equivalent to the skin state of a human being about 60 years old) to have young state change within 15 days under the condition that the concentration is 2000-50000 cells/mL, and can remarkably delay skin aging after 30 days of treatment, maintain the water-oil balance of the skin and increase the skin elasticity. The freeze-dried powder has obvious effect on the aspect of anti-aging improvement of aged skin.
The invention also provides an anti-aging human umbilical vein endothelial cell solution, which is prepared by mixing oligopeptide stock solution serving as a solvent and the freeze-dried powder serving as a solute according to the technical scheme. In the invention, the solution preferably contains 2000-50000 personal umbilical vein endothelial cells per milliliter, and more preferably 50000 personal umbilical vein endothelial cells per milliliter. In the present invention, the raw material for preparing the oligopeptide stock solution preferably comprises the following components in percentage by mass (%): water 70, propylene glycol 1, hydrolyzed collagen 3, acetylcysteine 2, acetyl-tyrosine 1, alanine 1, aloe extract 2, witch hazel extract 2, scutellaria extract 2, sophora flavescens root extract 2, glycyrrhiza inflata root extract 2, oligopeptide-12, betaine 2, p- hydroxyacetophenone 1, 1, 2-hexanediol 1, allantoin 1, sodium hyaluronate 2, dipotassium glycyrrhizinate 1, and oat peptide 2 (the oligopeptide stock solution is made by Shanghai posture Bo Biotech Co., Ltd., product number: O-0001). The oligopeptide stock solution disclosed by the invention has no harm to the skin, is beneficial to the generation of collagen and elastin, and maintains the young elasticity of the skin. Because the oligopeptide is a micro molecular structure, the oligopeptide can easily permeate through epidermal cells and directly reach the dermis layer below the epidermal cells and can be quickly absorbed by the dermis cells, so that the oligopeptide used as the lyophilized powder solvent of the endothelial cells of the umbilical veins of the human umbilical cord can promote the subsequent absorption of the lyophilized powder of the cells to directly reach the dermis layer. The invention selects 14-month-old mice, and the D-galactose is injected subcutaneously on the back to induce the subacute aging model of the mice, so as to construct an animal model with aged skin. The lyophilized powder of human umbilical vein endothelial cells is dissolved by oligopeptide stock solution to prepare the lyophilized powder of stem cells, and the lyophilized powder of stem cells is applied to the skin on the back of a mouse, so that the gloss of aged skin can be improved, the thickness of the skin can be increased, the number of subcutaneous blood vessels can be reduced, the water content of the skin can be increased, and/or the oil content of the skin can be reduced, and the skin can be changed from rough to fine.
The application of the human umbilical vein endothelial cells in preparing the anti-aging product, the anti-aging freeze-dried powder and the anti-aging solution are further described in detail with reference to the following specific examples, and the technical scheme of the invention includes but is not limited to the following examples.
Example 1
Separation and culture of clinical-grade primary human umbilical vein endothelial cells
TABLE 1 consumable List
TABLE 2 list of reagents
Separation, preparation and passage amplification of clinical-grade primary human umbilical vein endothelial cells
1 before the experiment, the experiment environment, instruments and equipment running conditions are checked. Wiping and disinfecting the operation table top of the biological safety cabinet by 75% ethanol solution, and wiping and disinfecting instruments and supplies by high pressure or alcohol.
2 ligating two ends of the umbilical cord tissue, soaking in a physiological saline bottle containing double antibodies, and transporting to a laboratory at the temperature of 2-8 ℃. Opening the bottle in a biological safety cabinet, discarding the preservation solution in the bottle, adding physiological saline (only by immersing the umbilical cord) into the bottle, and washing by shaking.
3 after discarding the saline, the umbilical cord was poured into a sterile 100mm petri dish and the surface of the umbilical cord was sprayed with a 75% ethanol solution. And cleaning the sprayed umbilical cord with normal saline for 2-3 times, adding the normal saline into a culture dish, dividing the umbilical cord into 3-4 cm sections with sterile scissors, clamping the center of the umbilical cord with forceps, pushing the umbilical cord towards two ends, and washing off blood.
4, putting the cleaned umbilical cord into a sterile 100mm culture dish containing normal saline, finding the umbilical vein, carrying out blunt peeling off the umbilical cord from the side of the vein, removing the vein, putting the umbilical cord into the culture dish, separating umbilical cord mesenchyme (Fahrenheit gelatin), and putting the umbilical cord mesenchyme (Fahrenheit gelatin) into the culture dish. The blood vessels in the petri dishes were flushed three more times with saline and a 5mL syringe, respectively, to remove the intravascular blood.
5 clamping one end of the blood vessel by a vascular clamp, injecting collagenase by a 5ml syringe until the blood vessel is obviously swelled, and clamping the other end by the vascular clamp. Cover the petri dish and put 5% CO2And digesting for 15min in a cell culture box at 37 ℃. The digested blood vessels were injected with physiological saline and 5mLThe container is washed for 3 times, the washed liquid is collected to a 50mL centrifuge tube and centrifuged for 5min at 1500 rpm.
6 discard the supernatant to add 10mL of complete ECM medium to the pellet to resuspend and make a cell suspension. And planting one hole of a six-hole plate according to a blood vessel of 4-5 cm. The medium was observed under a microscope and changed every 3 days.
7 when the cell fusion degree reaches 80%, adding 5mL of physiological saline to wash out residual culture medium in the bottle. After the physiological saline is discarded, 1mL of Tryple-Express is added, and the mixture is kept stand and digested for about 2min at room temperature or 37 ℃ until the cells are observed to be transparent and round by a microscope and are gently shaken and floated.
8, adding 5mL of physiological saline to stop digestion after digestion, washing residual cells on the lower bottle wall, transferring the liquid to a centrifuge tube, and centrifuging for 5min at 1500 rpm. The supernatant was discarded and the six well plates were inoculated in a ratio of 1: 3. Placing into an incubator under 5% CO2And culturing at 37 ℃.
9 observing the cell state in the six-hole plate cell culture bottle by using a microscope, and carrying out passage when the cell fusion degree reaches 80%. Adding 10mL of physiological saline to wash off residual culture medium in the bottle, discarding the physiological saline, adding 1mL of Tryple-Express, standing and digesting for 2min at room temperature or 37 ℃, adding 5mL of physiological saline into digested cells to stop digestion, washing residual cells on the wall of the bottle, transferring the liquid to a centrifuge tube, and centrifuging for 5min at 1500 rpm.
10 discard the supernatant, add 20mL of complete medium to the pellet and resuspend to make a cell suspension. After counting 20. mu.L, the cells were inoculated into a flask at a rate of 100 ten thousand/T-175 for amplification culture. Placing into an incubator under 5% CO2And culturing at 37 ℃.
Identification and purity detection of clinical grade human umbilical vein endothelial cells (results are shown in figure 1)
Identification of clinical grade human umbilical vein endothelial cells
1 spreading the prepared and amplified cells on a 24-well plate, washing the cells with DPBS for 3 times when the cell fusion degree is 80%, fixing the cells with 4% PFA for 15min, and washing the cells with DPBS for 3 times.
2 Primary antibody (Anti-Von Willebrand Factor antibody, cat # ab154193, abcam) was prepared in a ratio of 1:500 using 5% BSA + 0.4% Triton X-100 permeabilization solution and incubated overnight at 4 ℃.
Removing primary antibody, washing with DPBS for 3 times, and preparing secondary antibody (Anti-rabbitIgG (H + L), F (ab')2Fragment (Alexa) with the permeabilizing solution at a ratio of 1:500
488Conjugate), cat No. 4412, Cell Signaling Technology, inc.) and the secondary antibody were incubated at room temperature for 1 h.
The secondary antibody was discarded 4 times, washed with DPBS 3 times, incubated with DAPI (cat. No. D9542, Sigma) at room temperature for 15min, and the staining solution of DAPI was discarded, washed with DPBS 3 times, observed with a fluorescence microscope (OLYMPUS IX51), and photographed.
5 Von Willebrand Factor (Von Willebrand Factor), a multimeric adhesion glycoprotein, synthesized and secreted by endothelial cells, is a key Factor involved in the process of angiogenesis, as shown in A in FIG. 1. The prepared cells were identified as endothelial cells by detecting positive expression of Von Willebrand Factor as 100% by immunofluorescence staining.
Purity detection of clinical grade human umbilical vein endothelial cells
1 spreading the prepared and amplified cells on a 6-hole plate (2 holes), washing residual culture medium in a bottle by using physiological saline until the cell fusion degree is 80%, discarding the physiological saline, adding 1mL of Tryple-Express, standing and digesting at room temperature or 37 ℃ for 2min, adding 5mL of physiological saline into the digested cells to stop digestion, and centrifuging for 100g for 5 min.
2 staining of cells PE Mouse Anti-Human CD31 (cat # 555446, BD Co.) and detecting the positive expression of CD31 by flow cytometry (cat # FACSCANTO II, BD Co.).
3 as shown in B in FIG. 1, endothelial cell adhesion molecule-1 (Platlet/endothelial cell adhesion molecule-1, PECAM-1 or CD31) is highly expressed at the endothelial cell-cell junction and acts as an adhesion stress response protein, which can maintain the integrity of endothelial cell junction and rapidly restore the vascular permeability barrier after inflammation or thrombosis. The expression rate of CD31 in the cells was 99.9% by detecting cell surface antigen, and the prepared cells were identified as endothelial cells with a purity of 99.9%.
Example 2
Preparation of clinical grade human umbilical vein endothelial cell freeze-dried powder
1. Taking P5 and amplifying clinical grade human umbilical vein endothelial cells qualified by quality identification, harvesting 1000 ten thousand of cells, re-suspending in 10mL of physiological saline, and placing in a refrigerator at-80 ℃ to freeze into a solid; thawing the cells, adding double distilled water with 10 times of volume to dilute the cell suspension, and then putting the cell suspension into a refrigerator with the temperature of 80 ℃ below zero for overnight freezing (repeating the step for 3 times) to obtain a cell suspension subjected to repeated freezing and thawing;
2. starting a vacuum freeze dryer, and freezing the cell suspension to-45 ℃; then starting a freeze dryer cold trap to refrigerate to-45 ℃, and vacuumizing to 20 Pa; opening the box body and heating to-20 deg.C, and maintaining for 240 min; continuously heating to 1 deg.C and maintaining for 240 min; continuously heating to 30 ℃ and preserving heat for 300 min; discharging at 36 ℃ after the detection is qualified; and (6) filling, inspecting and warehousing.
Example 3
Effect of clinical grade lyophilized powder of human umbilical vein endothelial cells on aging skin of mouse
Experimental procedure
Female Kunming mice of 9 months old were purchased from Witonglihua, raised to 12 months old, weighed, depilated on the back and subcutaneously injected with D-galactose (cat # D8310, Beijing Soilebao Tech. Co., Ltd.) at a dose of 1mg/g for 30 days, once a day, for skin aging and molding.
One week after the end of D-galactose injection, mice were randomized into 4 groups: 1) control (oligopeptide stock solution); 2)2000 cells/mL; 3)10000 cells/mL; 3)50000cells/mL for different groups of treatments (all lyophilized cells were dissolved in oligopeptide stock solution). 0.1mL of clinical grade human umbilical vein endothelial cell freeze-dried powder with different concentrations is smeared on the back of the mouse every day until the freeze-dried powder is absorbed.
The body weight of the mouse is measured once a week, the skin aging condition of the mouse is recorded by taking a picture every 15 days, the water content, the oil content and the elasticity of the back skin of the mouse are detected by a skin detector after the treatment is finished, and the anti-skin aging effect of the freeze-dried powder is evaluated.
The results are analyzed as shown in fig. 2 (change of mouse weight in 4 weeks after clinical grade human umbilical vein endothelial cell lyophilized powder intervention), as shown in fig. 2, the mouse weight is not significantly changed in four weeks after intervention treatment, which indicates that the mouse is in a normal diet state, and the weight is not significantly increased and has a small part of descending trend due to aging of the mouse.
As shown in figure 3 (clinical grade human umbilical vein endothelial cell lyophilized powder anti-mouse skin aging effect result; cell lyophilized powder is divided into three concentration gradients of 2000cells/mL, 10000cells/mL and 50000cells/mL (dissolved in oligopeptide stock solution, intervention period is four weeks)), at day0, the back skin of 4 groups of aging mice becomes thin, the skin color is dark, and subcutaneous blood vessels are obvious. After 15 days of clinical grade human umbilical vein endothelial cell freeze-dried powder treatment, compared with a control group, mice of three treatment groups with different concentrations have bright skin color, and the skin is thickened and subcutaneous blood vessels are obviously reduced by visual observation. After one month of treatment, the skin state of the mice of the cell freeze-dried powder treatment groups with three concentrations is still remarkably improved compared with that of the control group on the same day, and the skin state of the mice of the treatment group with 50000cells/mL is better.
TABLE 3 clinical grade lyophilized powder of human umbilical vein endothelial cell for resisting skin aging of mice-moisture, oil and elasticity of skin
Note: within a certain range, the water content is high, the oil content is low, the skin elasticity index is good, and the skin is proved to be more youthful; the higher the oil content and the lower the water content, the coarser the skin is.
After 4 weeks of treatment, the skin conditions of the mice treated by different groups are quantitatively evaluated, and the water content, oil content and elasticity of the skin of the mice are detected by a skin monitor. As can be seen from Table 3, the lyophilized human umbilical vein endothelial cell powder is dose-dependent in its anti-aging effect on mouse skin. Along with the increase of the concentration of the cell freeze-dried powder, the water content of the skin of the mouse is increased, the oil content is reduced, and the skin state is changed from rough to delicate.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. Application of human umbilical vein endothelial cells in preparing antiaging product is provided.
2. Application of human umbilical vein endothelial cells in preparing products for resisting skin aging is provided.
3. Application of human umbilical vein endothelial cells in preparing products for resisting skin aging caused by D-galactose.
4. Application of human umbilical vein endothelial cells in preparing a product for improving the luster of aged skin.
5. Application of human umbilical vein endothelial cells in preparing products for increasing thickness of aged skin.
6. The application of human umbilical vein endothelial cells in preparing a product for reducing the number of subcutaneous blood vessels of aged skin.
7. Use of human umbilical vein endothelial cells for the manufacture of a product for increasing the water content and/or reducing the oil content of aged skin.
8. Application of human umbilical vein endothelial cells in preparing a product for improving the roughness of aged skin.
9. An anti-aging human umbilical vein endothelial cell freeze-dried powder, wherein the preparation method of the freeze-dried powder comprises the following steps:
and (3) repeatedly freezing and thawing the human umbilical vein endothelial cells, and freeze-drying in vacuum to obtain the human umbilical vein endothelial cell freeze-dried powder.
10. An anti-aging lyophilized human umbilical vein endothelial cell powder solution, which is prepared by mixing oligopeptide stock solution as a solvent and the lyophilized powder of claim 9 as a solute.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110911754.2A CN113750118A (en) | 2021-08-10 | 2021-08-10 | Application of human umbilical vein endothelial cells in preparation of anti-aging product, anti-aging freeze-dried powder and anti-aging solution |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110911754.2A CN113750118A (en) | 2021-08-10 | 2021-08-10 | Application of human umbilical vein endothelial cells in preparation of anti-aging product, anti-aging freeze-dried powder and anti-aging solution |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113750118A true CN113750118A (en) | 2021-12-07 |
Family
ID=78788885
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110911754.2A Pending CN113750118A (en) | 2021-08-10 | 2021-08-10 | Application of human umbilical vein endothelial cells in preparation of anti-aging product, anti-aging freeze-dried powder and anti-aging solution |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113750118A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116098899A (en) * | 2022-12-16 | 2023-05-12 | 中国科学技术大学 | Anti-aging preparation and application thereof |
-
2021
- 2021-08-10 CN CN202110911754.2A patent/CN113750118A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 赖红梅: "医疗损害责任纠纷研究", 武汉大学出版社, pages: 256 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116098899A (en) * | 2022-12-16 | 2023-05-12 | 中国科学技术大学 | Anti-aging preparation and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106109496B (en) | Human umbilical cord mesenchymal stem cells extract freeze-drying powder and preparation method | |
| CN103536967B (en) | A kind of method for removing cells preparing extracellular matrix support material | |
| CN106982821A (en) | Umbilical cord mesenchymal stem cells clinic freezes protection liquid composition and application thereof | |
| CN106821938A (en) | A kind of preparation method of human mesenchymal stem cell freeze-dried powder | |
| CN104195098A (en) | Dendrobium officinale stem cell and isolated culture method thereof | |
| CN108159078A (en) | A kind of Porcine HGF freeze-dried powder, preparation method and application | |
| CN101802177A (en) | Matter stem cell and extract the method for its secretory product between extracting from the human or animal embryo | |
| CN105920042A (en) | Umbilical cord mesenchymal stem cell injection with anti-aging function and preparation method thereof | |
| CN104955464A (en) | Compositions and methods for treating and preventing tissue injury and disease | |
| CN104622902B (en) | It is a kind of for treating the stem cell medicine of liver fibrosis | |
| CN105820998A (en) | Isolation extraction and culture method for human adipose-derived stem cells (ADSCs) for clinical back-transfusion grade cell therapy | |
| CN104814980A (en) | Production method and applications of human embryo fibroblasts | |
| CN108753708A (en) | A kind of preparation method of Stem Cell Activity factor freeze-dried powder | |
| CN110448572A (en) | A kind of preparation method of umbilical cord mesenchymal stem cells active matter and the compound of cord blood stem cell active matter | |
| CN106367388A (en) | Method for protecting umbilical cord mesenchymal stem cell culture supernatant at 4 DEG C | |
| CN104523753A (en) | Preparation method, product and application of human umbilical cord mesenchymal stem cell cultural supernatant active factor and cell lysis buffer | |
| CN110193025B (en) | Exosome freeze-dried powder derived from human adipose-derived stem cells, and preparation method and application thereof | |
| CN109453200A (en) | The preparation method of mostly tissue-derived mescenchymal stem cell factor lytic freeze-dried powder | |
| CN104399125A (en) | Method for differentiating epidermal stem cells to sweat gland-like epithelial cells | |
| CN104173252B (en) | A kind of biological beauty preparation of the cell containing autologous substrate | |
| CN113069532A (en) | Skin whitening and anti-aging medicine, health product or cosmetic containing exosome | |
| CN106924285A (en) | A kind of placenta mesenchyma stem cell parenteral solution and its preparation method and application | |
| CN113750118A (en) | Application of human umbilical vein endothelial cells in preparation of anti-aging product, anti-aging freeze-dried powder and anti-aging solution | |
| CN114392276A (en) | Anti-aging medicine prepared from improved anti-aging umbilical cord stem cells and related cosmetic products | |
| CN108066750A (en) | Stem cell and its secretion are used to treat the new application of skin burn |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20211207 |