CN107158035A - Treatment bedsore external-use gel containing high concentration human mesenchymal stem cell excretion body extract and preparation method thereof - Google Patents

Treatment bedsore external-use gel containing high concentration human mesenchymal stem cell excretion body extract and preparation method thereof Download PDF

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Publication number
CN107158035A
CN107158035A CN201710346752.7A CN201710346752A CN107158035A CN 107158035 A CN107158035 A CN 107158035A CN 201710346752 A CN201710346752 A CN 201710346752A CN 107158035 A CN107158035 A CN 107158035A
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cell
stem cell
mesenchymal stem
human mesenchymal
excretion body
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张冰晶
鲁振宇
韩洪起
秦臻
熊亮
高婧彧
徐悦
黄文敬
闻婧
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Tianjin Puri Purcell Biotechnology Co Ltd
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Tianjin Puri Purcell Biotechnology Co Ltd
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    • 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
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived 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
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • 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
    • A61K9/0014—Skin, i.e. galenical aspects of topical compositions
    • 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
    • A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
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    • 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
    • C12N5/0603—Embryonic cells ; Embryoid bodies
    • C12N5/0605—Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
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    • 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
    • C12N5/0662—Stem cells
    • C12N5/0663—Bone marrow mesenchymal stem cells (BM-MSC)
    • 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
    • C12N5/0662—Stem cells
    • C12N5/0664—Dental pulp stem cells, Dental follicle stem cells
    • 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
    • C12N5/0662—Stem cells
    • C12N5/0665—Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood
    • 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
    • C12N5/0662—Stem cells
    • C12N5/0667—Adipose-derived stem cells [ADSC]; Adipose stromal stem cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • 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
    • C12N5/0662—Stem cells
    • C12N5/0668—Mesenchymal stem cells from other natural sources
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/70—Undefined extracts
    • C12N2500/80—Undefined extracts from animals
    • C12N2500/84—Undefined extracts from animals from mammals

Abstract

The invention discloses a kind for the treatment of bedsore external-use gel containing high concentration human mesenchymal stem cell excretion body extract and preparation method thereof, human mesenchymal stem cell can secrete a variety of substantial amounts of cell factors in incubation, the deep enough skin base layer of these cell factors energy promotes the growth of skin histology differentiation, angiogenesis and granulation tissue, promote the structural remodeling and Regeneration and Repair of injured cutaneous tissue, wound healing.These cell factors and immunologic active material can adjust microenvironment, reduce bacterium infection and the inflammatory reaction of injury tissue.Wherein human mesenchymal stem cell secretion contains the compositions such as Multiple electrolytes injection to be extracted after repeating hungry culture.Reparation and treatment available for bedsore caused by a variety of causes.

Description

Treatment bedsore external application containing high concentration human mesenchymal stem cell excretion body extract is coagulated Glue and preparation method thereof
Technical field
The present invention relates to a kind of external-use gel preparation and preparation method thereof, especially containing high concentration human mesenchymal stem cell Treatment bedsore external-use gel of excretion body extract and preparation method thereof.
Background technology
Mescenchymal stem cell (mesenchymal stem cells, MSC) is the one group of foreign cell for coming from matrix group, can To organize to obtain from most of human body.They have self-renewal capacity, tissue repair, immunoregulation capability, can be to middle embryo Layer lineage, for example：Fat cell, osteocyte, cartilage cell etc., additionally can to and other germ-layer lineage cells point Change, for example to epidermal cell vascular endothelial cell break up.
Excretion body (exosome) is small vesica, diameter about 20-140nm, under physiology or pathologic condition, by a variety of The multivesicular body that intercellular membrane (en-dosomalmembrane) is formed in the mode of sprouting, participates in intercellular information interchange, and join With into a variety of pathological processes.Not only there is its specific proteins molecule from the excretion body of different tissues, and Also comprising the key molecule that it functions.The excretion body that existing research is proved mescenchymal stem cell secretion is reducing cardiac muscle damage Hinder scope, the damage of protection acute tubular, promote to play an important role in terms of nerve regneration, reduction injury of lungs.It is in immune tune Also played an important role in terms of section and vascularization.Therefore the reparation tool for being applied to skin injury has very important significance.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of containing the extraction of high concentration human mesenchymal stem cell excretion body Treatment bedsore external-use gel of thing and preparation method thereof, is carried using the excretion body of stem cell secretion for disease of skin such as treatment bedsores Medicine safely, effectively, easy to use is supplied.
In order to solve the above-mentioned technical problem, the technical solution adopted by the present invention is：One kind is done containing high concentration human mesenchyme The preparation method of the treatment bedsore external-use gel of cell excretion body extract, comprises the following steps：
(1) human mesenchymal stem cell culture：P2-P6 human mesenchymal stem cell recovery, inoculation batch are chosen from cell bank Secondary to be designated as first batch, second lot, the 3rd batch, inoculum density is respectively 50%-60%, 35%- of maximum degrees of fusion 45%th, 20%-30%；It without phenol red DMEM/DF12 and volume by volume concentration is 10% tire ox that complete medium used in culture cell, which is, Serum；Cell inoculation is completed, and is put in CO2gas incubator culture, 37 DEG C of temperature, carbon dioxide volume by volume concentration 5%；
(2) when first batch cell fusion degree reaches 70%-80%, cell is cleaned 2 times with physiological saline, is electrolysed with compound Matter parenteral solution carries out hungry culture；
(3) after starvation is cultivated 12-24 hours, first batch cell fusion degree reaches 90%-95%, collects in cell culture Clearly, 5-8min is centrifuged under the conditions of 3000-4500g and removes precipitation, take supernatant after centrifugation standby；
(4) the second lot cell of culture is taken, when cell fusion degree reaches 70%-80%, second is cleaned with physiological saline Batch cell 2 times, hungry culture is carried out with cells and supernatant in step (3)；
(5) after starvation is cultivated 12-24 hours, second lot cell fusion degree reaches 90%-95%, collects in cell culture Clearly, 5-8min is centrifuged under the conditions of 3000-4500g and removes precipitation, take supernatant after centrifugation standby；
(6) the 3rd batch cell of culture is taken, when cell fusion degree reaches 70%-80%, the 3rd is cleaned with physiological saline Batch cell 2 times, hungry culture is carried out with cells and supernatant in step (5)；
(7) after starvation is cultivated 12-24 hours, the 3rd batch cell degrees of fusion reaches 90%-95%, collects in cell culture Clearly, 5-8min is centrifuged under the conditions of 3000-4500g and removes precipitation, take supernatant after centrifugation standby；
(8) the step of collecting after continuous hungry three times (7) supernatant extracts therein dry thin with 0.22um membrane filtrations It is extracellular to secrete body, it is resuspended with the compound electrolyte solution of 0.5-2 times of step (7) supernatant volume and separates the excretion body extract solution obtained, It is put in 4 DEG C of refrigerators and preserves stand-by；
(9) gel basic components：Contain medicinal macromolecule stabilizer 2.5-10g, NMF 5- in per 100ml base gels 10ml, skin penetration enhancer 5-10ml, Multiple electrolytes injection surplus, mixing are 6.0-8.0 with rear adjustment pH value；
(10) by obtained excretion body extract solution in step (8) and the base gel of step (9) preparation according to volume ratio 1： 0.5-1：2 ratio is well mixed, and is filled in sterile cillin bottle, is preserved stand-by in 4 DEG C of refrigerators, and the bedsore that obtains medical treatment external application is coagulated Glue.
The human mesenchymal stem cell is human umbilical cord mesenchymal stem cells, fat mesenchymal stem cell, medulla mesenchyma are done Cell, placenta mesenchyma stem cell, menstrual blood mescenchymal stem cell or dental pulp mescenchymal stem cell.
The medicinal macromolecule stabilizer is one kind in sodium alginate, Sodium Hyaluronate, chitosan and HES.
The NMF is propane diols or glycerine.
The Multiple electrolytes injection is the compound electrolyte glucose MG3 notes that Otsuka Pharmaceutical (China) Co., Ltd. produces Penetrate liquid.
The skin penetration enhancer is dimethyl sulfoxide (DMSO).
Made from above-mentioned preparation method outside the treatment bedsore containing high concentration human mesenchymal stem cell excretion body extract Use gel.
The beneficial effects of the invention are as follows：Mescenchymal stem cell excretion body will be arrived and be prepared into gel preparations so that cell is secreted Excretion body preserved and utilized, and the time of survival can be can reach and be promoted epidermal cell to increase for a long time with longer, The healing time of wound can be shortened, and play and diminish inflammation, promote the effects such as ulcer healing for not recovering for a long time.
Brief description of the drawings
Fig. 1 is the mescenchymal stem cell figure of the method culture of the present invention；
Fig. 2 is that different hungry each cytokine secretion amounts of number of times compare figure.
Embodiment
The present invention is further described with specific embodiment below in conjunction with the accompanying drawings, and its specific embodiment is construed as By way of example only, it is not limited, it is impossible to limit protection scope of the present invention with following illustrations.
The preparation of the treatment bedsore external-use gel containing high concentration human mesenchymal stem cell excretion body extract of the present invention Method, comprises the following steps：
(1) human mesenchymal stem cell culture：P2-P6 human mesenchymal stem cell recovery, inoculation batch are chosen from cell bank Secondary to be designated as first batch, second lot, the 3rd batch, inoculum density is respectively 50%-60%, 35%- of maximum degrees of fusion 45%th, 20%-30%；It without phenol red DMEM/DF12 and volume by volume concentration is 10% tire ox that complete medium used in culture cell, which is, Serum；Cell inoculation is completed, and is put in CO2gas incubator culture, 37 DEG C of temperature, carbon dioxide volume by volume concentration 5%；
(2) when first batch cell fusion degree reaches 70%-80%, cell is cleaned 2 times with physiological saline, is electrolysed with compound Matter parenteral solution carries out hungry culture；
(3) after starvation is cultivated 12-24 hours, first batch cell fusion degree reaches 90%-95%, collects in cell culture Clearly, 5-8min is centrifuged under the conditions of 3000-4500g and removes precipitation, take supernatant after centrifugation standby；
(4) the second lot cell of culture is taken, when cell fusion degree reaches 70%-80%, second is cleaned with physiological saline Batch cell 2 times, hungry culture is carried out with cells and supernatant in step (3)；
(5) after starvation is cultivated 12-24 hours, second lot cell fusion degree reaches 90%-95%, collects in cell culture Clearly, 5-8min is centrifuged under the conditions of 3000-4500g and removes precipitation, take supernatant after centrifugation standby；
(6) the 3rd batch cell of culture is taken, when cell fusion degree reaches 70%-80%, the 3rd is cleaned with physiological saline Batch cell 2 times, hungry culture is carried out with cells and supernatant in step (5)；
(7) after starvation is cultivated 12-24 hours, the 3rd batch cell degrees of fusion reaches 90%-95%, collects in cell culture Clearly, 5-8min is centrifuged under the conditions of 3000-4500g and removes precipitation, take supernatant after centrifugation standby；
(8) the step of collecting after continuous hungry three times (7) supernatant extracts therein dry thin with 0.22um membrane filtrations It is extracellular to secrete body, it is resuspended with the compound electrolyte solution of 0.5-2 times of step (7) supernatant volume and separates the excretion body extract solution obtained, It is put in 4 DEG C of refrigerators and preserves stand-by；
(9) gel basic components：Contain medicinal macromolecule stabilizer 2.5-10g, NMF 5- in per 100ml base gels 10ml, skin penetration enhancer 5-10ml, Multiple electrolytes injection surplus, mixing are 6.0-8.0 with rear adjustment pH value；
(10) by obtained excretion body extract solution in step (8) and the base gel of step (9) preparation according to volume ratio 1： 0.5-1：2 ratio is well mixed, and is filled in sterile cillin bottle, is preserved stand-by in 4 DEG C of refrigerators, and the bedsore that obtains medical treatment external application is coagulated Glue.
The human mesenchymal stem cell is human umbilical cord mesenchymal stem cells, fat mesenchymal stem cell, medulla mesenchyma are done Cell, placenta mesenchyma stem cell, menstrual blood mescenchymal stem cell or dental pulp mescenchymal stem cell.
The medicinal macromolecule stabilizer is one kind in sodium alginate, Sodium Hyaluronate, chitosan and HES.
The NMF is propane diols or glycerine.
The Multiple electrolytes injection is the compound electrolyte glucose MG3 notes that Otsuka Pharmaceutical (China) Co., Ltd. produces Penetrate liquid.
The skin penetration enhancer is dimethyl sulfoxide (DMSO).
Made from above-mentioned preparation method outside the treatment bedsore containing high concentration human mesenchymal stem cell excretion body extract Use gel.
Mescenchymal stem cell of the present invention can secrete a variety of substantial amounts of excretion bodies, the function of these excretion bodies in incubation It is relevant with its derived cell, therefore the excretion body of human mesenchymal stem cell secretion can promote skin histology differentiation, angiogenesis And the growth of granulation tissue, so as to promote the structural remodeling and Regeneration and Repair of injured cutaneous tissue, reduce the production of scar tissue It is raw；Adjustable local organization microenvironment, promotes angiogenesis, improves local blood supply, so as to reach alleviation ischemia symptom, promotion group Knit the effect of new life；The immunologic function of local organization is can also adjust, bacterium infection and the inflammatory reaction of wound tissue is reduced.This hair The bright method using continuous hungry mescenchymal stem cell, within a short period of time with regard to human mesenchyme's extract of high concentration can be obtained, A large amount of stem cell excretion bodies are therefrom isolated by excretion body extractor, wherein in human mesenchymal stem cell excretion external use gel Contain sodium alginate and Multiple electrolytes injection as stabilizer, and containing NMF, skin penetration enhancer, form it into The spawn of stable uniform.Treatment for the skin injury disease such as bedsore.
Embodiment 1
(1) human umbilical cord mesenchymal stem cells culture：P4 human umbilical cord mesenchymal stem cells recovery is chosen from cell bank, carefully Born of the same parents' sum 8.4*107.Inoculation batch is designated as first batch, second lot, the 3rd batch, and inoculum density is respectively maximum degrees of fusion 60% (every bottle of inoculating cell number：9*106, six bottles of inoculating cell), 40% (every bottle of inoculating cell number：6*106, inoculating cell four Bottle), 20% (inoculating cell number：3*106, two bottles of inoculating cell), cultivating system is 40ml.Cultivated completely used in culture cell It without phenol red DMEM/DF12 and volume by volume concentration is 10% hyclone that base, which is, and inoculating cell bottle is T-175 blake bottles.Cell connects Plant and complete, be put in CO2gas incubator culture, 37 DEG C of temperature, carbon dioxide volume by volume concentration 5%；
(2) cultivate 30 hours, when now first batch cell fusion degree reaches 75%, see (Fig. 1), cleaned with physiological saline Cell 2 times, every bottle with 40ml Multiple electrolytes injections (Otsuka Pharmaceutical (China) Co., Ltd. produce compound electrolyte glucose MG3 parenteral solutions) carry out hungry culture；
(3) after starvation is cultivated 18 hours, first batch cell fusion degree collects cells and supernatant, 4000g bars up to 95% Under part centrifuge 8min remove precipitation, take centrifugation after supernatant be put in 4 DEG C it is standby；
(4) the second lot cell of culture is taken, now cell fusion degree reaches 70%, second lot is cleaned with physiological saline Cell 2 times, every bottle carries out hungry culture with cells and supernatant 40ml in step (3)；
(5) after starvation is cultivated 18 hours, second lot cell fusion degree collects cells and supernatant, 4000g bars up to 95% Under part centrifuge 8min remove precipitation, take centrifugation after supernatant be put in 4 DEG C it is standby；
(6) the 3rd batch cell of culture is taken, now cell fusion degree reaches 70%, the 3rd batch is cleaned with physiological saline Cell 2 times, every bottle carries out hungry culture with cells and supernatant 40ml in step (5)；
(7) after starvation is cultivated 24 hours, the 3rd batch cell degrees of fusion collects cells and supernatant, 4000g bars up to 90% Under part centrifuge 8min remove precipitation, take centrifugation after supernatant be put in 4 DEG C it is standby.
(8) each 40ml of standby supernatant in step (3), step (5) and step (7) is used into excretion in cells and supernatant Body quick-speed extraction apparatus (Chinese application number 201720177150.9) secretes stem cell excretion body therein, and with 10ml physiological saline Excretion body is resuspended with 40ml compound electrolytes solution after cleaning 2 times, cytokine content therein is surveyed in the physical examination of ultrasonication excretion, See (Fig. 2).
(9) each 40ml of standby supernatant in step (3), step (5) and step (7) is secreted it using excretion body extractor In stem cell excretion body, and with 10 physiological saline clean 2 times after with 30ml compound electrolytes solution be resuspended excretion body be stored in In sterile cillin bottle, i.e. umbilical cord mesenchymal stem cells excretion body extract is put in 4 DEG C of refrigerators and preserves stand-by.
(10) prepared by gel solution：By proportioning, by medicinal macromolecule stabilizer sodium alginate 2.5g, sterile NMF third Glycol 5ml, skin penetration enhancer dimethyl sulfoxide (DMSO) 5ml, are dissolved in 90ml compound electrolyte solution, are configured to homogeneous gel Solution for later use；
(11) external-use gel preparation is prepared：Umbilical cord mesenchymal stem cells excretion body extract by volume：Gel solution= 1:1 ratio is mixed, and after well mixed, is dispensed into pre-charge injector, be put into 4 DEG C of refrigerators freeze it is stand-by.
(12) zoopery：The SD rats 50 of 7 week old are chosen, 10 groups are equally divided into, preparing rat by pressure sore method lacks Courageous and upright damage model, skin damaged area is the wound of 2 centimetres of diameter, and for 1. blank group, (wound smears compound electrolyte to experiment packet Solution)；2. negative control group (wound smear gel basic ingredient)；3. group I (smear hungry one day excretion body and prepare by wound Gel)；4. experimental group II (wound smears gel prepared by hungry two days excretion bodies)；5. experimental group III (smear hungry by wound Gel prepared by three days excretion bodies), smear secondary daily, each 0.2ml, respectively upon administration the 0th, 2,4,6,8,10 days it is right The diameter of wound is measured, and statistical damage area (table one).
Embodiment 2
(1) human adipose mesenchymal stem cells culture：P2 human adipose mesenchymal stem cells recovery is chosen from cell bank, carefully Born of the same parents' sum 8.4*107.Inoculation batch is designated as first batch, second lot, the 3rd batch, and inoculum density is respectively maximum degrees of fusion 60% (every bottle of inoculating cell number：9*106, six bottles of inoculating cell), 40% (every bottle of inoculating cell number：6*106, inoculating cell four Bottle), 20% (inoculating cell number：3*106, two bottles of inoculating cell), cultivating system is 40ml.Cultivated completely used in culture cell It without phenol red DMEM/DF12 and volume by volume concentration is 10% hyclone that base, which is, and inoculating cell bottle is T-175 blake bottles.Cell connects Plant and complete, be put in CO2gas incubator culture, 37 DEG C of temperature, carbon dioxide volume by volume concentration 5%；
(2) cultivate 24 hours, when now first batch cell fusion degree reaches 70%, cell cleaned 2 times with physiological saline, Every bottle carries out hungry culture with 40ml Multiple electrolytes injections；
(3) after starvation is cultivated 20 hours, first batch cell fusion degree collects cells and supernatant, 4000g bars up to 95% Under part centrifuge 8min remove precipitation, take centrifugation after supernatant be put in 4 DEG C it is standby；
(4) the second lot cell of culture is taken, now cell fusion degree reaches 70%, second lot is cleaned with physiological saline Cell 2 times, every bottle carries out hungry culture with cells and supernatant 40ml in step (3)；
(5) after starvation is cultivated 20 hours, second lot cell fusion degree collects cells and supernatant, 4000g bars up to 95% Under part centrifuge 8min remove precipitation, take centrifugation after supernatant be put in 4 DEG C it is standby；
(6) the 3rd batch cell of culture is taken, now cell fusion degree reaches 75%, the 3rd batch is cleaned with physiological saline Cell 2 times, every bottle carries out hungry culture with cells and supernatant 40ml in step (5)；
(7) after starvation is cultivated 20 hours, the 3rd batch cell degrees of fusion collects cells and supernatant, 4000g bars up to 95% Under part centrifuge 8min remove precipitation, take centrifugation after supernatant be put in 4 DEG C it is standby.
(8) each 40ml of standby supernatant in step (3), step (5) and step (7) is secreted it using excretion body extractor In stem cell excretion body, and excretion body be resuspended with 30ml compound electrolytes solution after being cleaned 2 times with 10ml physiological saline preserved In sterile cillin bottle, i.e. fat mesenchymal stem cell excretion body extract is put in 4 DEG C of refrigerators and preserves stand-by.
(10) prepared by gel solution：By proportioning, by medicinal macromolecule stabilizer sodium alginate 2.5g, sterile NMF is sweet Oily 5ml, skin penetration enhancer dimethyl sulfoxide (DMSO) 5ml, are dissolved in 90ml compound electrolyte solution, are configured to homogeneous gel molten Liquid is stand-by；
(11) external-use gel preparation is prepared：Fat mesenchymal stem cell excretion body extract by volume：Gel solution= 2:1 ratio is mixed, and after well mixed, is dispensed into pre-charge injector, be put into 4 DEG C of refrigerators freeze it is stand-by.
Table 1 is the data of zoopery
Group Blank group Negative group Group I Experimental group II Experimental group III
Number of animals 10 10 10 10 10
D0 wound diameters 2±0.1 2±0.1 2±0.1 2±0.1 2±0.1
D0T/C 1 1 1 1 1
D2 wound diameters 2±0.2 1.9±0.1 1.8±0.2 1.8±0.2 1.7±0.2
D2T/C 1 0.95 0.9 0.9 0.85
D4 wound diameters 1.8±0.2 1.6±0.1 1.4±0.2 1.2±0.2 1.1±0.1
D4T/C 1 0.89 0.78 0.67 0.61
D6 wound diameters 1.5±0.1 1.3±0.2 1.2±0.1 1.0±0.1 0.6±0.1
D6T/C 1 0.87 0.8 0.67 0.4
D8 wound diameters 1.2±0.1 1.0±0.1 0.9±0.1 0.8±0.1 0.3±0
D8T/C 1 0.83 0.75 0.67 0.25
D10 wound diameters 1.1±0.1 0.9±0.1 0.8±0.1 0.6±0.1 0.2±0
D10T/C 1 0.82 0.73 0.55 0.18
The wound diameter unit of table 1 for centimetre；T/C is the ratio of each group wound diameter and blank group wound diameter；Examined through Q Test, compared with blank control group, P<0.05.
In summary, present disclosure is not limited in the above embodiments, and the knowledgeable people in same area can Can propose other embodiments easily within the technological guidance's thought of the present invention, but this embodiment is included in this hair Within the scope of bright.

Claims (7)

1. a kind of preparation method of the treatment bedsore external-use gel containing high concentration human mesenchymal stem cell excretion body extract, its It is characterised by, comprises the following steps：
(1) human mesenchymal stem cell culture：P2-P6 human mesenchymal stem cell recovery, inoculation batch note are chosen from cell bank For first batch, second lot, the 3rd batch, inoculum density be respectively the 50%-60% of maximum degrees of fusion, 35%-45%, 20%-30%；It without phenol red DMEM/DF12 and volume by volume concentration is 10% hyclone that complete medium used in culture cell, which is,； Cell inoculation is completed, and is put in CO2gas incubator culture, 37 DEG C of temperature, carbon dioxide volume by volume concentration 5%；
(2) when first batch cell fusion degree reaches 70%-80%, cell is cleaned 2 times with physiological saline, noted with compound electrolyte Penetrate liquid and carry out hungry culture；
(3) after starvation is cultivated 12-24 hours, first batch cell fusion degree reaches 90%-95%, collects cells and supernatant, 5-8min is centrifuged under the conditions of 3000-4500g and removes precipitation, takes supernatant after centrifugation standby；
(4) the second lot cell of culture is taken, when cell fusion degree reaches 70%-80%, second lot is cleaned with physiological saline Cell 2 times, hungry culture is carried out with cells and supernatant in step (3)；
(5) after starvation is cultivated 12-24 hours, second lot cell fusion degree reaches 90%-95%, collects cells and supernatant, 5-8min is centrifuged under the conditions of 3000-4500g and removes precipitation, takes supernatant after centrifugation standby；
(6) the 3rd batch cell of culture is taken, when cell fusion degree reaches 70%-80%, the 3rd batch is cleaned with physiological saline Cell 2 times, hungry culture is carried out with cells and supernatant in step (5)；
(7) after starvation is cultivated 12-24 hours, the 3rd batch cell degrees of fusion reaches 90%-95%, collects cells and supernatant, 5-8min is centrifuged under the conditions of 3000-4500g and removes precipitation, takes supernatant after centrifugation standby；
(8) the step of collecting after continuous hungry three times (7) supernatant is extracted outside stem cell therein with 0.22um membrane filtrations Body is secreted, is resuspended with the compound electrolyte solution of 0.5-2 times of step (7) supernatant volume and separates the excretion body extract solution obtained, be put in 4 Preserve stand-by in DEG C refrigerator；
(9) gel basic components：Contain medicinal macromolecule stabilizer 2.5-10g, NMF 5- in per 100ml base gels 10ml, skin penetration enhancer 5-10ml, Multiple electrolytes injection surplus, mixing are 6.0-8.0 with rear adjustment pH value；
(10) by obtained excretion body extract solution in step (8) and the base gel of step (9) preparation according to volume ratio 1：0.5- 1：2 ratio is well mixed, and is filled in sterile cillin bottle, is preserved stand-by in 4 DEG C of refrigerators, the bedsore that obtains medical treatment external-use gel.
2. the treatment bedsore external-use gel according to claim 1 containing high concentration human mesenchymal stem cell excretion body extract Preparation method, it is characterised in that the human mesenchymal stem cell is that human umbilical cord mesenchymal stem cells, fat mesenchymal are dry thin Born of the same parents, mesenchymal stem cells MSCs, placenta mesenchyma stem cell, menstrual blood mescenchymal stem cell or dental pulp mescenchymal stem cell.
3. the treatment bedsore external-use gel according to claim 1 containing high concentration human mesenchymal stem cell excretion body extract Preparation method, it is characterised in that the medicinal macromolecule stabilizer be sodium alginate, Sodium Hyaluronate, chitosan and hydroxyl second One kind in base starch.
4. the treatment bedsore external-use gel according to claim 1 containing high concentration human mesenchymal stem cell excretion body extract Preparation method, it is characterised in that the NMF be propane diols or glycerine.
5. the treatment bedsore external-use gel according to claim 1 containing high concentration human mesenchymal stem cell excretion body extract Preparation method, it is characterised in that the Multiple electrolytes injection be Otsuka Pharmaceutical (China) Co., Ltd. produce compound electricity Solve matter glucose MG3 parenteral solutions.
6. the treatment bedsore external-use gel according to claim 1 containing high concentration human mesenchymal stem cell excretion body extract Preparation method, it is characterised in that the skin penetration enhancer be dimethyl sulfoxide (DMSO).
7. carried made from the preparation method as described in claim any one of 1-6 containing high concentration human mesenchymal stem cell excretion body Take the treatment bedsore external-use gel of thing.
CN201710346752.7A 2017-05-17 2017-05-17 Treatment bedsore external-use gel containing high concentration human mesenchymal stem cell excretion body extract and preparation method thereof Pending CN107158035A (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105412153A (en) * 2015-12-10 2016-03-23 中国人民解放军第二军医大学 Application of exosome excreted by mesenchymal stem cells for preparing medicine for preventing and treating hepatitis c virus
CN105477016A (en) * 2015-11-13 2016-04-13 中国人民解放军第二军医大学 Application of exosome derived from human mesenchymal stem cells to resistance to tissue fibrosis and scar forming
CN106474155A (en) * 2016-10-19 2017-03-08 天津普瑞赛尔生物科技有限公司 External-use gel preparation containing human umbilical cord mesenchymal stem cells extract and its production and use

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105477016A (en) * 2015-11-13 2016-04-13 中国人民解放军第二军医大学 Application of exosome derived from human mesenchymal stem cells to resistance to tissue fibrosis and scar forming
CN105412153A (en) * 2015-12-10 2016-03-23 中国人民解放军第二军医大学 Application of exosome excreted by mesenchymal stem cells for preparing medicine for preventing and treating hepatitis c virus
CN106474155A (en) * 2016-10-19 2017-03-08 天津普瑞赛尔生物科技有限公司 External-use gel preparation containing human umbilical cord mesenchymal stem cells extract and its production and use

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
遆冬冬 等: "炎症激活间充质干细胞源微泡对糖尿病慢性创面愈合的促进作用观察", 《解放军医学杂志》 *

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