CN111150746A - Stem cell repair liquid for treating gynecological inflammation by vaginal administration - Google Patents
Stem cell repair liquid for treating gynecological inflammation by vaginal administration Download PDFInfo
- Publication number
- CN111150746A CN111150746A CN202010010906.7A CN202010010906A CN111150746A CN 111150746 A CN111150746 A CN 111150746A CN 202010010906 A CN202010010906 A CN 202010010906A CN 111150746 A CN111150746 A CN 111150746A
- Authority
- CN
- China
- Prior art keywords
- stem cell
- solution
- parts
- suppository
- vagina
- 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
- 210000000130 stem cell Anatomy 0.000 title claims abstract description 32
- 230000008439 repair process Effects 0.000 title claims abstract description 20
- 239000007788 liquid Substances 0.000 title claims abstract description 18
- 206010061218 Inflammation Diseases 0.000 title abstract description 16
- 230000004054 inflammatory process Effects 0.000 title abstract description 16
- 210000004027 cell Anatomy 0.000 claims abstract description 23
- 239000011159 matrix material Substances 0.000 claims abstract description 16
- 102000029816 Collagenase Human genes 0.000 claims abstract description 4
- 108060005980 Collagenase Proteins 0.000 claims abstract description 4
- 229960002424 collagenase Drugs 0.000 claims abstract description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 claims abstract 7
- 239000000243 solution Substances 0.000 claims description 38
- 239000000829 suppository Substances 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 239000002609 medium Substances 0.000 claims description 14
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 claims description 12
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 12
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 12
- 229920001661 Chitosan Polymers 0.000 claims description 9
- 108010010803 Gelatin Proteins 0.000 claims description 9
- 239000008273 gelatin Substances 0.000 claims description 9
- 229920000159 gelatin Polymers 0.000 claims description 9
- 235000019322 gelatine Nutrition 0.000 claims description 9
- 235000011852 gelatine desserts Nutrition 0.000 claims description 9
- 239000008215 water for injection Substances 0.000 claims description 9
- 238000005303 weighing Methods 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 8
- 230000000694 effects Effects 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 claims description 6
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 5
- 210000002966 serum Anatomy 0.000 claims description 5
- 239000003102 growth factor Substances 0.000 claims description 4
- BQRXMLXVGQMIBO-UHFFFAOYSA-N 1,1-bis(sulfanyl)ethanol Chemical compound CC(O)(S)S BQRXMLXVGQMIBO-UHFFFAOYSA-N 0.000 claims description 3
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 claims description 3
- 229930182816 L-glutamine Natural products 0.000 claims description 3
- 230000010261 cell growth Effects 0.000 claims description 3
- 238000001816 cooling Methods 0.000 claims description 3
- 229940079593 drug Drugs 0.000 claims description 3
- 239000003797 essential amino acid Substances 0.000 claims description 3
- 235000020776 essential amino acid Nutrition 0.000 claims description 3
- 238000000465 moulding Methods 0.000 claims description 3
- 238000004806 packaging method and process Methods 0.000 claims description 3
- 238000001935 peptisation Methods 0.000 claims description 3
- 229920003023 plastic Polymers 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 229960005322 streptomycin Drugs 0.000 claims description 3
- 210000001789 adipocyte Anatomy 0.000 claims description 2
- 238000012258 culturing Methods 0.000 claims description 2
- 239000012530 fluid Substances 0.000 claims 3
- 230000001678 irradiating effect Effects 0.000 claims 1
- 238000002156 mixing Methods 0.000 claims 1
- 210000001215 vagina Anatomy 0.000 abstract description 12
- 210000001519 tissue Anatomy 0.000 abstract description 8
- 108091003079 Bovine Serum Albumin Proteins 0.000 abstract description 5
- 239000012091 fetal bovine serum Substances 0.000 abstract description 5
- 239000000463 material Substances 0.000 abstract description 5
- 230000008929 regeneration Effects 0.000 abstract description 5
- 238000011069 regeneration method Methods 0.000 abstract description 5
- 238000006731 degradation reaction Methods 0.000 abstract description 2
- 210000002615 epidermis Anatomy 0.000 abstract description 2
- 210000000981 epithelium Anatomy 0.000 abstract description 2
- 239000001963 growth medium Substances 0.000 abstract description 2
- 230000001737 promoting effect Effects 0.000 abstract description 2
- 238000000034 method Methods 0.000 description 8
- 206010046914 Vaginal infection Diseases 0.000 description 6
- 201000008100 Vaginitis Diseases 0.000 description 6
- 150000001413 amino acids Chemical group 0.000 description 6
- 230000000890 antigenic effect Effects 0.000 description 4
- 210000004877 mucosa Anatomy 0.000 description 4
- 241000701806 Human papillomavirus Species 0.000 description 3
- 241000186660 Lactobacillus Species 0.000 description 3
- 210000000577 adipose tissue Anatomy 0.000 description 3
- 239000004599 antimicrobial Substances 0.000 description 3
- 238000007689 inspection Methods 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 2
- 241000341655 Human papillomavirus type 16 Species 0.000 description 2
- 206010020751 Hypersensitivity Diseases 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 230000000845 anti-microbial effect Effects 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 230000000875 corresponding effect Effects 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000000857 drug effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 239000006210 lotion Substances 0.000 description 2
- 210000004400 mucous membrane Anatomy 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 206010006784 Burning sensation Diseases 0.000 description 1
- 208000036649 Dysbacteriosis Diseases 0.000 description 1
- 208000027244 Dysbiosis Diseases 0.000 description 1
- 208000004483 Dyspareunia Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 101710132594 Protein E6 Proteins 0.000 description 1
- 101710132595 Protein E7 Proteins 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 208000003251 Pruritus Diseases 0.000 description 1
- 208000007893 Salpingitis Diseases 0.000 description 1
- 206010046793 Uterine inflammation Diseases 0.000 description 1
- 206010051873 Vaginal relaxation Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 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
- 238000003287 bathing Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000003636 conditioned culture medium Substances 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 238000006471 dimerization reaction Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000007140 dysbiosis Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 210000004996 female reproductive system Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000007803 itching Effects 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000009469 supplementation Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 208000002003 vulvitis Diseases 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0653—Adipocytes; Adipose tissue
-
- 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
- A61K35/35—Fat tissue; Adipocytes; Stromal cells; Connective tissues
-
- 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
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0034—Urogenital system, e.g. vagina, uterus, cervix, penis, scrotum, urethra, bladder; Personal lubricants
-
- 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
- A61K9/02—Suppositories; Bougies; Bases therefor; Ovules
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/02—Drugs for genital or sexual disorders; Contraceptives for disorders of the vagina
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Reproductive Health (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Cell Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biotechnology (AREA)
- Gynecology & Obstetrics (AREA)
- Rheumatology (AREA)
- Endocrinology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Inorganic Chemistry (AREA)
- Pain & Pain Management (AREA)
- Microbiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Urology & Nephrology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Developmental Biology & Embryology (AREA)
- Immunology (AREA)
- Virology (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to a stem cell repair liquid for vagina administration to treat gynecological inflammation, which is a stem cell extract for vagina wall, wherein the stem cell extract is prepared by digesting tissue cells by collagenase and subculturing the tissue cells in a DMEM culture medium containing FBS (fetal bovine serum) with low sugar DMEM, and the culture condition is 5% CO2In, temperature settingCulturing at 37 deg.C for 2 days for passage, collecting cells for passage 3-5, and performing subsequent treatment. The invention combines stem cell regeneration with material with good biocompatibility, is placed on the inner wall of the vagina, and in the gradual degradation process of the material, the cells continue to proliferate and secrete matrix to finally form corresponding tissues, thereby achieving the purposes of promoting the regeneration of the epithelium on the vaginal wall, repairing the damaged inner wall of the vagina and recovering the moistening of the epidermis on the inner wall of the vagina.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to a stem cell repair liquid for treating gynecological inflammation by vaginal administration.
Background
The inflammation of female reproductive system is gynecological inflammation, including vulvitis, vaginitis, metritis, salpingitis, etc. Gynecological inflammation is a common and frequently encountered disease of women. And in severe cases may even be carcinogenic. Various inflammations are mostly caused by damage to the wall of organs due to external factors, such as vaginitis. At present, the common methods for clinically treating gynecological inflammation are local supplementation of bioengineering lactobacillus, use of antibiotic antimicrobial preparations and the like, but the methods have limitations. Bioengineered lactobacilli have the potential to pose new biological risks and not all lactobacilli have an antibacterial effect. Antibiotic antimicrobial agents, while inhibiting the growth of pathogenic microorganisms, also kill normal flora, leading to further dysbacteriosis. Moreover, the use of broad spectrum antibiotics also presents a range of other problems such as infection with resistant strains of bacteria.
At present, traditional Chinese medicine compositions are adopted to treat gynecological inflammation, but most of the traditional Chinese medicine external lotion has weakened drug effect after being diluted by lotion water, and the drug can not flow out conveniently, which is not as good as the drug effect of an antimicrobial preparation. Antimicrobial agents are associated with allergic reactions, and have local irritation, itching and burning sensations, which are not ideal. The gel preparation is convenient to use, good in tolerance of patients, low in allergy rate and easy to form a functional isolation membrane, so that the gel preparation is widely concerned.
The basic method is that autologous normal tissue cells cultured and expanded in vitro, such as vaginal wall epithelial cells, mucosal cells and the like, are adsorbed on a material with good biocompatibility and placed on the position of the inner wall of a vagina, and in the gradual degradation process of the material, the cells continue to proliferate and secrete matrix to finally form corresponding tissues, so that the aims of promoting the regeneration of the vaginal wall epithelium, repairing the damaged inner wall of the vagina and recovering the moistening of the epidermis of the inner wall of the vagina are fulfilled.
Disclosure of Invention
The invention aims to provide a stem cell repair liquid for treating gynecological inflammation by vaginal administration, which can effectively promote the regeneration and repair of vaginal inner wall injury.
A stem cell repairing liquid for vagina administration to treat gynecological inflammation is a stem cell extract for vagina wall, and is prepared by digesting tissue cells with collagenase and subculturing in DMEM medium.
Preferably, the DMEM medium contains the following substances in parts by weight: 8-10 parts of low-sugar DMEM medium, 2-3 parts of serum, 2-3 parts of L-glutamine, 2-3 parts of dimercaptoethanol, 1-2 parts of non-essential amino acid, 1-3 parts of cell growth factors EGF and bFGF, 1-2 parts of streptomycin double-antibody solution and 2-6 parts of LIF.
Preferably, the serum is FBS.
Preferably, the EGF activity unit is 75U/ml and the bFGF activity unit is 100U/ml.
Preferably, the culture conditions are at 5% CO2In (1), the temperature was set to 37 ℃.
Preferably, the cells are cultured once for 2 days and passaged, and the cells are collected for the subsequent treatment at passage 3-5.
Preferably, the stem cell repairing solution is filled in a suppository shape made of a degradable material.
Preferably, the tissue cells are adipocytes of the body.
The stem cell-containing repair liquid is prepared by the following specific method:
1) weighing 5.0-6.0 g of gelatin, adding into 5-7 mL of water for injection, stirring in a water bath at 80 ℃ to dissolve to obtain a gelatin matrix solution, and keeping the temperature at 40-50 ℃ for later use.
2) Weighing 0.5-1.0 g of polyvinyl alcohol, adding the polyvinyl alcohol into 5-6mL of water for injection, placing the mixture in a water bath at 90 ℃ after the mixture is naturally swelled to be heated and dissolved to prepare a polyvinyl alcohol solution, and keeping the temperature at 40-50 ℃ for later use.
3) Weighing 0.5-0.6 g of chitosan, dissolving in 3-4 mL of water for injection, and preparing a chitosan solution for later use after natural peptization.
4) Under the aseptic condition, the polyvinyl alcohol solution and the chitosan solution are uniformly mixed, added into the gelatin matrix solution and uniformly stirred to prepare the suppository matrix solution, and the suppository matrix solution is kept at the temperature of 40-50 ℃ for later use.
5) Adding 5-6ml of cultured stem cell repairing solution into the suppository matrix solution under aseptic condition, stirring, injecting into a mold for molding, cooling and solidifying to obtain suppository, sterilizing for 30 min by ultraviolet irradiation, and packaging in plastic package. Each suppository is about 3g in weight and contains about 5ng of basic fibroblast growth factor.
The formula of the invention enables the repairing liquid to activate DC cells at female private parts by the technical principle of arranging amino acid sequence numbers, and after the DC cells are activated, the efficacy of the repairing liquid can penetrate mucous membranes to reach 100 percent of penetration, thereby killing deeply infected viruses. The polypeptide protein in the formula repairs the whole vaginal mucosa. In the whole repair process, along with the continuous and powerful overall function, other gynecological inflammations of the female disappear.
A homodimeric protein of two identical amino acid chains, each amino acid chain comprising: (1) a signal peptide, (2) a targeting unit, (3) a dimerization motif, and (4) an antigenic unit, the targeting unit comprising a peptide that is homologous to seq id no: 1 and the antigenic unit comprises an amino acid sequence of a Human Papilloma Virus (HPV), such as an antigenic unit comprising the amino acid sequence of HPV16 and/or HPV18, such as an antigenic unit derived from the early protein E6 and/or E7 of HPV16 and/or HPV 18. The amino acid sequence number is arranged to activate the low DC cell in the formula by a technical means, and when the DC cell is activated, the repair liquid can penetrate through a mucous membrane to kill deeply infected viruses. The polypeptide protein in the formula repairs the whole vaginal mucosa. In the whole repair process, along with the continuous and powerful overall function, other gynecological inflammations of the female disappear. The whole formula generates a synergistic effect.
The invention has the beneficial effects that: the stem cell regeneration and repair is adopted to effectively treat gynecological inflammation such as vaginitis, the medicine is put into a human body for a period of time and then automatically degrades through the administration form of the suppository, a stem cell repair liquid release channel is opened, and the vaginal mucosa is helped to recover the self-repairing capacity.
Detailed Description
It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The stem cell repairing liquid for treating gynecological inflammation through vaginal administration is prepared by the steps of culturing adipose tissues in vitro, digesting the adipose tissues by using a DMEM (modified eagle medium) containing collagenase, filtering, subculturing by using a DMEM medium containing 10% Fetal Bovine Serum (FBS) by mass, subculturing by using a serum-free DMEM medium, subculturing once in 2 days, collecting cells for passage 3-5 generations, and performing subsequent treatment; digesting cells by using a DMEM medium containing EDTA, washing by using a PBS solution, re-suspending by using a serum-free DMEM medium, ultrasonically cracking and breaking the cells, and centrifuging a broken product to obtain a supernatant; the supernatant was filtered through a filter having a pore size of 5000 angstroms. The culture medium comprises the following components in parts by weight: 9 parts of low-sugar DMEM medium, 3 parts of serum, 2 parts of L-glutamine, 2 parts of dimercaptoethanol, 1 part of non-essential amino acid, 2 parts of cell growth factors EGF and bFGF (wherein the activity unit of the EGF is 75U/ml, and the activity unit of the bFGF is 100U/ml), 1 part of streptomycin double antibody solution and 5 parts of LIF.
The stem cell-containing repair liquid is prepared by the following specific method:
1) weighing 5.0-6.0 g of gelatin, adding into 5-7 mL of water for injection, stirring in a water bath at 80 ℃ to dissolve to obtain a gelatin matrix solution, and keeping the temperature at 40-50 ℃ for later use.
2) Weighing 0.5-1.0 g of polyvinyl alcohol, adding the polyvinyl alcohol into 5-6mL of water for injection, placing the mixture in a water bath at 90 ℃ after the mixture is naturally swelled to be heated and dissolved to prepare a polyvinyl alcohol solution, and keeping the temperature at 40-50 ℃ for later use.
3) Weighing 0.5-0.6 g of chitosan, dissolving in 3-4 mL of water for injection, and preparing a chitosan solution for later use after natural peptization.
4) Under the aseptic condition, the polyvinyl alcohol solution and the chitosan solution are uniformly mixed, added into the gelatin matrix solution and uniformly stirred to prepare the suppository matrix solution, and the suppository matrix solution is kept at the temperature of 40-50 ℃ for later use.
5) Adding 5-6ml of cultured stem cell repairing solution into the suppository matrix solution under aseptic condition, stirring, injecting into a mold for molding, cooling and solidifying to obtain suppository, sterilizing for 30 min by ultraviolet irradiation, and packaging in plastic package. Each suppository is about 3g in weight and contains about 5ng of basic fibroblast growth factor.
The basic fibroblast growth factor can be determined by the prior art, such as enzyme-linked immunosorbent assay (ELISA) for determining the content of the basic fibroblast growth factor in the conditioned medium. Firstly, a microporous plate is coated by a purified antibody to prepare a solid phase antibody. Adding corresponding antigen into micropores coated with the monoclonal antibody, combining, washing, adding enzyme-labeled anti-antibody, washing, and developing with color developing agent. The shade of the color is positively correlated with the growth factor content in the sample. The absorbance (OD value) was measured at a wavelength of 450nm with a microplate reader, and the sample concentration was calculated. And pure serum-free culture solution was used as a control. As a result of the measurement, the adipose tissue-derived culture broth of this example contained 1.285. + -. 0.506) ng/ml of basic fibroblast growth factor.
The stem cell repairing liquid is prepared into suppository, the suppository is put into a human body for a period of time and then is degraded automatically, a release channel of the stem cell repairing liquid is opened, and the vaginal mucosa is helped to recover the self-repairing capability.
The application effect experiment for treating vaginitis:
30 vaginitis patients are recruited, the age is 25-45 years, and the using method is as follows: the subject directly uses the stem cell repairing liquid of the invention, and the using method comprises the following steps: it is administered once a month after bathing every night. The suppository is inserted into deep vagina, vagina observation and leucorrhea routine inspection are carried out on the same day after the start of the experiment, inspection reports and observation results are kept, inspection is carried out after one month, and all results of 30 patients with vaginitis restore the tightness, the elasticity and the damaged tissues to be repaired. The symptoms of vaginal relaxation, dyspareunia and the like of a patient are improved after 1 month of use.
The above description is only a preferred embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (6)
1. A vaginal drug for treating gynecological inflammationThe stem cell repairing solution is a stem cell extract used for the vaginal wall, and is characterized in that the stem cell extract is prepared by digesting tissue cells by collagenase and subculturing the tissue cells in a DMEM medium, wherein the DMEM medium is a low-sugar DMEM medium, and the culture condition is that the tissue cells are subjected to subculturing by 5% CO2And (3) setting the temperature to be 37 ℃, culturing for 2 days for one passage, collecting cells for passage 3-5 passages for subsequent treatment, wherein the DMEM medium comprises the following components in parts by weight: 8-10 parts of low-sugar DMEM medium, 2-3 parts of serum, 2-3 parts of L-glutamine, 2-3 parts of dimercaptoethanol, 1-2 parts of non-essential amino acid, 1-3 parts of cell growth factors EGF and bFGF, 1-2 parts of streptomycin double-antibody solution and 2-6 parts of LIF.
2. The stem cell repair fluid according to claim 1, wherein the serum is FBS.
3. The stem cell repair fluid of claim 2, wherein the EGF activity unit is 75U/ml and the bFGF activity unit is 100U/ml.
4. The stem cell repair solution according to claim 3, wherein the stem cell repair solution is prepared into a suppository.
5. The stem cell repair fluid according to claim 4, wherein the tissue cells are adipocytes of the body.
6. The stem cell repairing liquid according to claim 5, wherein the preparation method of the suppository containing the stem cell repairing liquid comprises the following steps:
1) weighing 5.0-6.0 g of gelatin, adding into 5-7 mL of water for injection, stirring in a water bath at 80 ℃ to dissolve to obtain a gelatin matrix solution, and keeping the temperature at 40-50 ℃ for later use;
2) weighing 0.5-1.0 g of polyvinyl alcohol, adding the polyvinyl alcohol into 5-6mL of water for injection, placing the mixture in a water bath at 90 ℃ after the mixture is naturally swelled to be heated and dissolved to prepare a polyvinyl alcohol solution, and keeping the temperature at 40-50 ℃ for later use;
3) weighing 0.5-0.6 g of chitosan, dissolving in 3-4 mL of water for injection, and preparing a chitosan solution for later use after natural peptization;
4) under the aseptic condition, uniformly mixing the polyvinyl alcohol solution and the chitosan solution, adding the mixture into the gelatin matrix solution, uniformly stirring to prepare a suppository matrix solution, and keeping the temperature at 40-50 ℃ for later use;
5) under aseptic conditions, adding 5-6ml of cultured stem cell repairing solution into the suppository matrix solution, stirring uniformly, injecting into a mold for molding, cooling and solidifying to obtain a suppository, irradiating with ultraviolet rays for sterilizing for 30 minutes, and packaging in a plastic package to obtain the suppository; each suppository is about 3g in weight and contains about 5ng of basic fibroblast growth factor.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010010906.7A CN111150746A (en) | 2020-01-06 | 2020-01-06 | Stem cell repair liquid for treating gynecological inflammation by vaginal administration |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010010906.7A CN111150746A (en) | 2020-01-06 | 2020-01-06 | Stem cell repair liquid for treating gynecological inflammation by vaginal administration |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111150746A true CN111150746A (en) | 2020-05-15 |
Family
ID=70561484
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010010906.7A Pending CN111150746A (en) | 2020-01-06 | 2020-01-06 | Stem cell repair liquid for treating gynecological inflammation by vaginal administration |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111150746A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113730329A (en) * | 2021-08-25 | 2021-12-03 | 杭州美约健康管理有限公司 | Autologous mucosal cell regeneration and repair composition, preparation method and application thereof |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110217385A1 (en) * | 2008-10-10 | 2011-09-08 | Team Youn Biomedical Technology Co., Ltd. | Method for extracting mesenchymal stem cell from human or animal embryo and for extracting the secretion product thereof |
| CN102600456A (en) * | 2012-03-22 | 2012-07-25 | 兰州大学第二医院 | Bioadhesive vaginal plug for treating vaginitis and cervicitis as well as preparation method of bioadhesive vaginal plug |
| CN103255103A (en) * | 2013-04-17 | 2013-08-21 | 广州爱菲科生物科技有限公司 | Serum-free adipose tissue-derived mesenchymal stem cell culture medium |
| US20150118748A1 (en) * | 2012-06-26 | 2015-04-30 | K-Stemcell Co., Ltd. | High-concentration stem cell production method |
| CN108524601A (en) * | 2018-05-22 | 2018-09-14 | 北京恒峰昊瑞生物科技有限公司 | A kind of vaginal jellies and preparation method thereof containing pleiotrophic factor |
| CN108992662A (en) * | 2018-08-23 | 2018-12-14 | 杭州元研细胞生物科技有限公司 | A kind of vagina of the factor containing mescenchymal stem cell is spraying and preparation method thereof |
| US20190046573A1 (en) * | 2015-09-07 | 2019-02-14 | Eduardo ALVARO GALUE | Process for obtaining a sprinkling compound of microvascular endothelial skin cells and mesenchymal stem cells and method of application for tissue regeneration |
| CN109439617A (en) * | 2018-12-25 | 2019-03-08 | 成都赋智健康科技有限公司 | A kind of stem cell serum-free culture medium and preparation method thereof |
| CN110628695A (en) * | 2019-11-12 | 2019-12-31 | 安徽科门生物科技有限公司 | Culture medium for stem cell amplification |
-
2020
- 2020-01-06 CN CN202010010906.7A patent/CN111150746A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110217385A1 (en) * | 2008-10-10 | 2011-09-08 | Team Youn Biomedical Technology Co., Ltd. | Method for extracting mesenchymal stem cell from human or animal embryo and for extracting the secretion product thereof |
| CN102600456A (en) * | 2012-03-22 | 2012-07-25 | 兰州大学第二医院 | Bioadhesive vaginal plug for treating vaginitis and cervicitis as well as preparation method of bioadhesive vaginal plug |
| US20150118748A1 (en) * | 2012-06-26 | 2015-04-30 | K-Stemcell Co., Ltd. | High-concentration stem cell production method |
| CN103255103A (en) * | 2013-04-17 | 2013-08-21 | 广州爱菲科生物科技有限公司 | Serum-free adipose tissue-derived mesenchymal stem cell culture medium |
| US20190046573A1 (en) * | 2015-09-07 | 2019-02-14 | Eduardo ALVARO GALUE | Process for obtaining a sprinkling compound of microvascular endothelial skin cells and mesenchymal stem cells and method of application for tissue regeneration |
| CN108524601A (en) * | 2018-05-22 | 2018-09-14 | 北京恒峰昊瑞生物科技有限公司 | A kind of vaginal jellies and preparation method thereof containing pleiotrophic factor |
| CN108992662A (en) * | 2018-08-23 | 2018-12-14 | 杭州元研细胞生物科技有限公司 | A kind of vagina of the factor containing mescenchymal stem cell is spraying and preparation method thereof |
| CN109439617A (en) * | 2018-12-25 | 2019-03-08 | 成都赋智健康科技有限公司 | A kind of stem cell serum-free culture medium and preparation method thereof |
| CN110628695A (en) * | 2019-11-12 | 2019-12-31 | 安徽科门生物科技有限公司 | Culture medium for stem cell amplification |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113730329A (en) * | 2021-08-25 | 2021-12-03 | 杭州美约健康管理有限公司 | Autologous mucosal cell regeneration and repair composition, preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR101595108B1 (en) | Nativetelopeptide Placental Collagen Compositions | |
| WO2008002064A1 (en) | Soft tissue filler composition for injection and preparation method thereof | |
| WO2016168950A9 (en) | Method for in vitro constructing salivary gland organoids and acinus-like | |
| AU2007265862A1 (en) | Soft tissue filler composition comprising autologous dermis-derived cell culture product and hyaluronic acid | |
| JP2020142114A (en) | Extracellular matrix compositions | |
| TW201343195A (en) | Cosmetic or skin regeneration promoting agent comprising supernatant of non-human dental pulp stem cell culture, and ion introducing method of protein | |
| CN106511387A (en) | Preparing method and application of chicken embryo extracts | |
| CN110638834A (en) | Stem cell exosome composite preparation for improving premature ovarian failure and application thereof | |
| Ding et al. | Galectin-1-induced skeletal muscle cell differentiation of mesenchymal stem cells seeded on an acellular dermal matrix improves injured anal sphincter | |
| Zhang et al. | Decellularized extracellular matrix and mesenchymal stem cells promote recovery in traumatic brain injury by synergistically enhancing neurogenesis and attenuating neuroinflammation | |
| CN111150746A (en) | Stem cell repair liquid for treating gynecological inflammation by vaginal administration | |
| CN113846064A (en) | FGF18 gene modified mesenchymal stem cell and preparation method and application thereof | |
| US20030202965A1 (en) | Methods and compositions for the preparation of cell transplants | |
| CN102172337B (en) | Tissue engineering skin with sebaceous gland-like structure and preparation method thereof | |
| KR20210021165A (en) | The pharmaceutical composition for wound healing comprising conditioned medium containing cell spheroid derived from adipose-derived stem cell | |
| US20220233601A1 (en) | Composition comprising exosomes derived from induced pluripotent stem cell-derived mesenchymal stem cell progenitor for prevention or treatment of non-alcoholic steatohepatitis | |
| CN108066749A (en) | Purposes of the Stem Cell Activity factor in skin injury drug | |
| CN108066748A (en) | New application of the excretion body in skin injury | |
| CN108066824A (en) | A kind of new method for preparing skin blemish medicine | |
| CN106562994A (en) | Preparation method for preparing duck embryos and application of duck embryos | |
| CN116421710A (en) | Collagen-stem cell preparation for promoting repair of skin wounds of diabetic pets | |
| CN110526958B (en) | Activity repair peptide Tvigour A and application thereof | |
| KR20160000030A (en) | Pharmaceutical adjuvant composition for treating damages of skin or blood vessel tissue | |
| KR102286779B1 (en) | Endometrial complex for treating endometrial damage, manufacturing method of endometrial complex and endometrial complex using the same | |
| CN106562995A (en) | Preparation method for preparing swan embryos and application of swan embryos |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20200515 |