CN113662906A - Preparation method of stem cell exosome anti-aging essence - Google Patents
Preparation method of stem cell exosome anti-aging essence Download PDFInfo
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
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Abstract
The invention discloses a preparation method of stem cell exosome anti-aging essence, which has the technical scheme main points that: the method comprises the following steps: s11, preparing materials: preparing the following raw materials in parts by weight: 2-10 parts of stem cell exosome suspension, 5-10 parts of dendrobium officinale extract, 2-5 parts of ganoderma lucidum fermentation filtrate, 1-3 parts of vitamin complex, 0.5-0.8 part of schizophyllan, 1-3 parts of towel gourd extract, 1-3 parts of plant polysaccharide and 20-65 parts of deionized water: s12, primary mixing: sequentially placing the dendrobium officinale extracting solution, the lucid ganoderma fermentation filtrate, the towel gourd extracting solution, the plant polysaccharide and the deionized water into a reaction kettle, controlling the rotating speed of the reaction kettle at 200-500r/min, and continuously mixing for 20-40min to obtain a mixed solution; has the advantages of no side effect, effective improvement of skin, aging delay, wrinkle elimination, damaged skin repair, and inflammation and scar inhibition.
Description
Technical Field
The invention belongs to the field of cosmetics, and particularly relates to a preparation method of stem cell exosome anti-aging essence.
Background
The stem cell exosome is a micro vesicle which can be secreted by most stem cells, has the diameter of about 30-150nm, and consists of lipid, protein and nucleic acid. Once the exosomes are released from stem cells by exocytosis, complex mixed factors can be transmitted to other cells as signal molecules, thereby changing the functions of other cells, and the exosomes are wrapped in a hard double-layer membrane which protects the contents of the exosomes and enables the exosomes to move in tissues for a long distance. The stem cell exosome is a tool for cell-free cell therapy due to the effects of proliferation, migration, regeneration, inflammation, scar control and the like of epithelial tissues, and the essence is one of facial skin care products and contains precious functional components.
Reference can be made to a Chinese patent with publication number CN108670913A, which discloses a plant essence with hair-blacking effect, relating to the technical field of daily chemical industry. The essence can be prepared from 0.3-1% of traditional Chinese medicine extract, 0.02-0.08% of preservative and 0.4-1% of vinasse hydrolysate containing amino acid nitrogen in percentage by weight. The traditional Chinese medicine components added in the invention comprise angelica, ligusticum wallichii, radix angelicae, mulberry, saffron and cacumen biotae, and the traditional Chinese medicine components are respectively extracted according to a specific extraction process and are mixed according to a specific proportion.
The above patent has the advantage of significant hair-blacking effect, but it also has drawbacks, such as: it cannot delay aging and improve skin quality, and is not suitable for all skin.
Disclosure of Invention
The invention aims to provide a preparation method of stem cell exosome anti-aging essence, and aims to solve the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme:
a preparation method of stem cell exosome anti-aging essence comprises the following steps:
s11, preparing materials: preparing the following raw materials in parts by weight: 2-10 parts of stem cell exosome suspension, 5-10 parts of dendrobium officinale extract, 2-5 parts of ganoderma lucidum fermentation filtrate, 1-3 parts of vitamin complex, 0.5-0.8 part of schizophyllan, 1-3 parts of towel gourd extract, 1-3 parts of plant polysaccharide and 20-65 parts of deionized water:
s12, primary mixing: sequentially placing the dendrobium officinale extracting solution, the lucid ganoderma fermentation filtrate, the towel gourd extracting solution, the plant polysaccharide and the deionized water into a reaction kettle, controlling the rotating speed of the reaction kettle at 200-500r/min and the temperature at 40-65 ℃, and continuously mixing for 20-40min to obtain a mixed solution;
s13, secondary mixing: sequentially adding stem cell exosome suspension, multivitamins and schizophyllan into the mixed solution obtained in S12, controlling the temperature of the reaction kettle at 30-40 ℃, continuously mixing for 1-4h, and taking out to obtain the essence;
s14, sterilizing and filling: and (5) sterilizing the essence obtained in the step (S13), and filling the sterilized essence by using filling equipment, wherein the net content of each bottle is controlled to be 50-100 ml.
Preferably, the suspension of the stem cell exosomes contains 20-35mg of protein per ml of solution.
Preferably, the preparation method of the dendrobium officinale extract comprises the following steps:
s21, selecting fresh and clean dendrobium officinale to dry, and then putting the dried dendrobium officinale into a crusher to be crushed to obtain dendrobium officinale powder;
s22, adding deionized water into the reaction kettle, heating to 50-70 ℃, adding the dendrobium officinale powder obtained in the S21, controlling the rotating speed of the reaction kettle to be 600r/min, continuously stirring for 20-50min to obtain a mixed solution, filtering the mixed solution to remove residual dendrobium officinale particles, adding a complex enzyme into the mixed solution for enzymolysis, and controlling the enzymolysis time to be 1-5 h;
s23, carrying out adsorption and elution operations on the mixed solution after enzymolysis to obtain a semi-finished product solution, and finally concentrating the semi-finished product solution to control the concentration of the semi-finished product solution after concentration to be 18-35Brix, thus obtaining the dendrobium officinale extracting solution.
Preferably, in S23, the mixed solution is subjected to an adsorption operation using a molecular sieve, and the mixed solution is subjected to an elution operation using an aqueous ethanol solution.
Preferably, the sterilization operation comprises the following specific steps:
s31, injecting the primary essence liquid into a sterilized glass container, and sealing the glass container with a sealing cover;
s32, starting an ultraviolet sterilizer to sterilize the outside of the glass container and the sealing cover, opening the sealing cover, sending the glass container and the primary essence liquid into the sterilization equipment, and pumping out air in the sterilization equipment by using a vacuum pump to ensure that the inside of the sterilization equipment is in a vacuum state;
s33, starting the sterilization equipment to sterilize the primary essence liquid in the glass container, and simultaneously mixing the primary essence liquid by using the stirring mechanism in the sterilization equipment.
Preferably, in S33, the stirring speed of the stirring mechanism is controlled to be 50-80r/min, and then the sterilization time of the sterilization equipment is controlled to be 3-5 h.
Preferably, the vitamin complex is formed by combining vitamin A, vitamin B6, vitamin E and vitamin H, wherein the mass ratio of the vitamin A to the vitamin B6 to the vitamin E to the vitamin H is 1: 1: 2: 1.5.
preferably, the plant polysaccharide is formed by combining flammulina velutipes polysaccharide, tremella polysaccharide, acronychia pedunculata polysaccharide, ginseng polysaccharide and cistanche polysaccharide, and the mass ratio of the flammulina velutipes polysaccharide to the tremella polysaccharide to the acronychia pedunculata polysaccharide to the ginseng polysaccharide to the cistanche polysaccharide is 2: 3: 2: 1: 2.
preferably, the stem cell exosome suspension is a mesenchymal stem cell exosome suspension, and the stem cell exosome suspension contains a skin-improving plasmid vector.
Preferably, the preparation method of the stem cell exosome suspension comprises the following steps:
s41, culturing the stem cells by using a serum-containing culture medium, extracting after the culture is finished to obtain the stem cells, and then placing the stem cells in the culture medium for culturing for 1-3 days;
s42, filtering to remove stem cells in the culture medium, filtering, putting the filtered stem cell solution into a centrifuge tube, centrifuging, removing supernatant in the centrifuge tube to obtain a stem cell suspension, and then precipitating;
s43, inoculating the precipitated stem cell suspension into a culture bottle at the cell density of 50000 cells/CM 2, and then culturing the culture bottle;
s44, after the culture is finished, collecting the stem cell suspension in all culture bottles into a centrifugal container, repeatedly centrifuging for 3-5 times, removing supernatant after each centrifugation, and finally obtaining a precipitation solution, namely the stem cell exosome suspension.
Compared with the prior art, the invention has the beneficial effects that:
when the preparation method of the stem cell exosome anti-aging essence is used, the stem cell exosome can promote the regeneration of fibroblasts and keratinocytes related to skin, and simultaneously secrete collagen, elastic fibrin and matrix components, so that the functions of repairing skin, promoting the angiogenesis required by the growth and nutrition supply of skin cells, inhibiting excessive inflammatory reaction and inhibiting scar formation are achieved;
the dendrobium officinale extracting solution contains natural moisture-preserving and antioxidant active ingredients such as polysaccharide, flavonoid, phenols and the like, can effectively resist the oxidation of the outside on the skin, thereby playing a very good skin-care effect, and in addition, the dendrobium officinale extracting solution is slightly cold in nature and has the effects of nourishing yin, moistening dryness, enriching the blood, tonifying qi and delaying senility;
the loofah extracting solution contains various saccharides, plant mucus, various vitamins and trace mineral substances, can maintain the normal water content of the horny layer of the skin of a human body, and has the advantages of slowing down dehydration, supplementing necessary moisture to the skin and keeping the skin tender and fine; meanwhile, the towel gourd extracting solution also contains a large amount of natural moisturizing factors, so that the skin is rich in elasticity and can be kept moist, the metabolism can be promoted, dirt in the deep layer of the skin can be removed, the generation of melanocytes can be inhibited, the rough skin can be improved, and wrinkles can be eliminated;
in conclusion, the invention adds the effects of beauty treatment and anti-aging through the supplement of the raw materials, can be suitable for any skin, and has the advantages of no side effect, effective improvement of the skin, aging delay, wrinkle elimination, damaged skin repair, inflammation inhibition and scar inhibition.
Drawings
FIG. 1 is a flow chart of the method of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Please refer to FIG. 1
Example 1
A preparation method of stem cell exosome anti-aging essence comprises the following steps:
s11, preparing materials: preparing the following raw materials in parts by weight: 6 parts of stem cell exosome suspension, 6 parts of dendrobium officinale extracting solution, 3 parts of lucid ganoderma fermentation filtrate, 2 parts of vitamin complex, 0.6 part of schizophyllan, 2 parts of towel gourd extracting solution, 2 parts of plant polysaccharide and 55 parts of deionized water:
s12, primary mixing: sequentially placing the dendrobium officinale extracting solution, the lucid ganoderma fermentation filtrate, the towel gourd extracting solution, the plant polysaccharide and deionized water into a reaction kettle, controlling the rotating speed of the reaction kettle at 300r/min and the temperature at 60 ℃, and continuously mixing for 30min to obtain a mixed solution;
s13, secondary mixing: sequentially adding stem cell exosome suspension, multivitamins and schizophyllan into the mixed solution obtained in S12, controlling the temperature of the reaction kettle at 37 ℃, continuously mixing for 2 hours, and taking out to obtain the essence;
s14, sterilizing and filling: and (5) sterilizing the essence obtained in the step (S13), and filling the sterilized essence by using filling equipment, wherein the net content of each bottle is controlled to be 50 ml.
In this embodiment, the stem cell exosome suspension preferably contains 35mg of protein per ml of solution.
In this embodiment, preferably, the preparation method of the dendrobium officinale extract solution is as follows:
s21, selecting fresh and clean dendrobium officinale to dry, and then putting the dried dendrobium officinale into a crusher to be crushed to obtain dendrobium officinale powder;
s22, adding deionized water into the reaction kettle, heating to 60 ℃, adding the dendrobium officinale powder obtained in the S21, controlling the rotating speed of the reaction kettle to be 600r/min, continuously stirring for 35min to obtain a mixed solution, filtering the mixed solution to remove residual dendrobium officinale particles, adding a compound enzyme into the mixed solution for enzymolysis, and controlling the enzymolysis time to be 3 hours;
s23, carrying out adsorption and elution operations on the mixed solution after enzymolysis to obtain a semi-finished product solution, and finally concentrating the semi-finished product solution to control the concentration of the semi-finished product solution after concentration to be 25Brix, thus obtaining the dendrobium officinale extracting solution.
In this example, in S23, the mixed solution is preferably subjected to an adsorption operation using a molecular sieve, and the mixed solution is preferably subjected to an elution operation using an aqueous ethanol solution.
In this embodiment, it is preferable that the sterilization operation includes the following steps:
s31, injecting the primary essence liquid into a sterilized glass container, and sealing the glass container with a sealing cover;
s32, starting an ultraviolet sterilizer to sterilize the outside of the glass container and the sealing cover, opening the sealing cover, sending the glass container and the primary essence liquid into the sterilization equipment, and pumping out air in the sterilization equipment by using a vacuum pump to ensure that the inside of the sterilization equipment is in a vacuum state;
s33, starting the sterilization equipment to sterilize the primary essence liquid in the glass container, and simultaneously mixing the primary essence liquid by using the stirring mechanism in the sterilization equipment.
In this embodiment, it is preferable that in S33, the stirring speed of the stirring mechanism is controlled to 60r/min, and then the sterilization time of the sterilization apparatus is controlled to 4 h.
In this embodiment, the vitamin complex is preferably a combination of vitamin a, vitamin B6, vitamin E, and vitamin H, wherein the mass ratio of vitamin a, vitamin B6, vitamin E, and vitamin H is 1: 1: 2: 1.5.
in this embodiment, preferably, the plant polysaccharide is a combination of flammulina velutipes polysaccharide, tremella polysaccharide, acronychia negra polysaccharide, ginseng polysaccharide and cistanche polysaccharide, and the mass ratio of the flammulina velutipes polysaccharide to the tremella fuciformis polysaccharide to the ginseng polysaccharide to the cistanche polysaccharide is 2: 3: 2: 1: 2.
in this embodiment, preferably, the stem cell exosome suspension is a mesenchymal stem cell exosome suspension, and the stem cell exosome suspension contains a skin-improving plasmid vector.
In this embodiment, preferably, the stem cell exosome suspension is prepared as follows:
s41, culturing the stem cells by using a serum-containing culture medium, extracting after the culture is finished to obtain the stem cells, and then placing the stem cells in the culture medium for 2 days;
s42, filtering to remove stem cells in the culture medium, filtering, putting the filtered stem cell solution into a centrifuge tube, centrifuging, removing supernatant in the centrifuge tube to obtain a stem cell suspension, and then precipitating;
s43, inoculating the precipitated stem cell suspension into a culture bottle at the cell density of 50000 cells/CM 2, and then culturing the culture bottle;
s44, after the culture is finished, collecting the stem cell suspension in all culture bottles into a centrifugal container, repeatedly centrifuging for 4 times, removing supernatant after each centrifugation, and finally obtaining a precipitation solution, namely the stem cell exosome suspension.
The invention has the beneficial effects that:
when the preparation method of the stem cell exosome anti-aging essence is used, the stem cell exosome can promote the regeneration of fibroblasts and keratinocytes related to skin, and simultaneously secrete collagen, elastic fibrin and matrix components, so that the functions of repairing skin, promoting the angiogenesis required by the growth and nutrition supply of skin cells, inhibiting excessive inflammatory reaction and inhibiting scar formation are achieved;
the dendrobium officinale extracting solution contains natural moisture-preserving and antioxidant active ingredients such as polysaccharide, flavonoid, phenols and the like, can effectively resist the oxidation of the outside on the skin, thereby playing a very good skin-care effect, and in addition, the dendrobium officinale extracting solution is slightly cold in nature and has the effects of nourishing yin, moistening dryness, enriching the blood, tonifying qi and delaying senility;
the loofah extracting solution contains various saccharides, plant mucus, various vitamins and trace mineral substances, can maintain the normal water content of the horny layer of the skin of a human body, and has the advantages of slowing down dehydration, supplementing necessary moisture to the skin and keeping the skin tender and fine; meanwhile, the towel gourd extracting solution also contains a large amount of natural moisturizing factors, so that the skin is rich in elasticity and can be kept moist, the metabolism can be promoted, dirt in the deep layer of the skin can be removed, the generation of melanocytes can be inhibited, the rough skin can be improved, and wrinkles can be eliminated;
in conclusion, the invention adds the effects of beauty treatment and anti-aging through the supplement of the raw materials, can be suitable for any skin, and has the advantages of no side effect, effective improvement of the skin, aging delay, wrinkle elimination, damaged skin repair, inflammation inhibition and scar inhibition.
Example 2
A preparation method of stem cell exosome anti-aging essence comprises the following steps:
s11, preparing materials: preparing the following raw materials in parts by weight: 2 parts of stem cell exosome suspension, 5 parts of dendrobium officinale extract, 2 parts of ganoderma lucidum fermentation filtrate, 1 part of vitamin complex, 0.5 part of schizophyllan, 1 part of towel gourd extract, 1 part of plant polysaccharide and 60 parts of deionized water:
s12, primary mixing: sequentially placing the dendrobium officinale extracting solution, the lucid ganoderma fermentation filtrate, the towel gourd extracting solution, the plant polysaccharide and deionized water into a reaction kettle, controlling the rotating speed of the reaction kettle at 200r/min and the temperature at 40 ℃, and continuously mixing for 20min to obtain a mixed solution;
s13, secondary mixing: sequentially adding stem cell exosome suspension, multivitamins and schizophyllan into the mixed solution obtained in S12, controlling the temperature of the reaction kettle at 30-40 ℃, continuously mixing for 1-4h, and taking out to obtain the essence;
s14, sterilizing and filling: and (5) sterilizing the essence obtained in the step (S13), and filling the sterilized essence by using filling equipment, wherein the net content of each bottle is controlled to be 50 ml.
In this embodiment, the stem cell exosome suspension preferably contains 20mg of protein per ml of solution.
In this embodiment, preferably, the preparation method of the dendrobium officinale extract solution is as follows:
s21, selecting fresh and clean dendrobium officinale to dry, and then putting the dried dendrobium officinale into a crusher to be crushed to obtain dendrobium officinale powder;
s22, adding deionized water into the reaction kettle, heating to 50 ℃, adding the dendrobium officinale powder obtained in the S21, controlling the rotating speed of the reaction kettle to be 600r/min, continuously stirring for 20min to obtain a mixed solution, filtering the mixed solution to remove residual dendrobium officinale particles, adding a compound enzyme into the mixed solution for enzymolysis, and controlling the enzymolysis time to be 1 h;
s23, carrying out adsorption and elution operations on the mixed solution after enzymolysis to obtain a semi-finished product solution, and finally concentrating the semi-finished product solution to control the concentration of the semi-finished product solution after concentration to be 18Brix, thus obtaining the dendrobium officinale extracting solution.
In this example, in S23, the mixed solution is preferably subjected to an adsorption operation using a molecular sieve, and the mixed solution is preferably subjected to an elution operation using an aqueous ethanol solution.
In this embodiment, it is preferable that the sterilization operation includes the following steps:
s31, injecting the primary essence liquid into a sterilized glass container, and sealing the glass container with a sealing cover;
s32, starting an ultraviolet sterilizer to sterilize the outside of the glass container and the sealing cover, opening the sealing cover, sending the glass container and the primary essence liquid into the sterilization equipment, and pumping out air in the sterilization equipment by using a vacuum pump to ensure that the inside of the sterilization equipment is in a vacuum state;
s33, starting the sterilization equipment to sterilize the primary essence liquid in the glass container, and simultaneously mixing the primary essence liquid by using the stirring mechanism in the sterilization equipment.
In this embodiment, it is preferable that in S33, the stirring speed of the stirring mechanism is controlled to 50r/min, and then the sterilization time of the sterilization apparatus is controlled to 3 hours.
In this embodiment, the vitamin complex is preferably a combination of vitamin a, vitamin B6, vitamin E, and vitamin H, wherein the mass ratio of vitamin a, vitamin B6, vitamin E, and vitamin H is 1: 1: 2: 1.5.
in this embodiment, preferably, the plant polysaccharide is a combination of flammulina velutipes polysaccharide, tremella polysaccharide, acronychia negra polysaccharide, ginseng polysaccharide and cistanche polysaccharide, and the mass ratio of the flammulina velutipes polysaccharide to the tremella fuciformis polysaccharide to the ginseng polysaccharide to the cistanche polysaccharide is 2: 3: 2: 1: 2.
in this embodiment, preferably, the stem cell exosome suspension is a mesenchymal stem cell exosome suspension, and the stem cell exosome suspension contains a skin-improving plasmid vector.
In this embodiment, preferably, the stem cell exosome suspension is prepared as follows:
s41, culturing the stem cells by using a serum-containing culture medium, extracting after the culture is finished to obtain the stem cells, and then placing the stem cells in the culture medium for 1 day;
s42, filtering to remove stem cells in the culture medium, filtering, putting the filtered stem cell solution into a centrifuge tube, centrifuging, removing supernatant in the centrifuge tube to obtain a stem cell suspension, and then precipitating;
s43, inoculating the precipitated stem cell suspension into a culture bottle at the cell density of 50000 cells/CM 2, and then culturing the culture bottle;
s44, after the culture is finished, collecting the stem cell suspension in all culture bottles into a centrifugal container, repeatedly centrifuging for 3 times, removing supernatant after each centrifugation, and finally obtaining a precipitation solution, namely the stem cell exosome suspension.
Example 3
A preparation method of stem cell exosome anti-aging essence comprises the following steps:
s11, preparing materials: preparing the following raw materials in parts by weight: 10 parts of stem cell exosome suspension, 10 parts of dendrobium officinale extract, 5 parts of ganoderma lucidum fermentation filtrate, 3 parts of vitamin complex, 0.8 part of schizophyllan, 3 parts of towel gourd extract, 3 parts of plant polysaccharide and 60 parts of deionized water:
s12, primary mixing: sequentially placing the dendrobium officinale extracting solution, the lucid ganoderma fermentation filtrate, the towel gourd extracting solution, the plant polysaccharide and deionized water into a reaction kettle, controlling the rotating speed of the reaction kettle at 500r/min and the temperature at 65 ℃, and continuously mixing for 40min to obtain a mixed solution;
s13, secondary mixing: sequentially adding stem cell exosome suspension, multivitamins and schizophyllan into the mixed solution obtained in S12, controlling the temperature of the reaction kettle at 40 ℃, continuously mixing for 4 hours, and taking out to obtain the essence;
s14, sterilizing and filling: and (5) sterilizing the essence obtained in the step (S13), and filling the sterilized essence by using filling equipment, wherein the net content of each bottle is controlled to be 100 ml.
In this embodiment, the stem cell exosome suspension preferably contains 35mg of protein per ml of solution.
In this embodiment, preferably, the preparation method of the dendrobium officinale extract solution is as follows:
s21, selecting fresh and clean dendrobium officinale to dry, and then putting the dried dendrobium officinale into a crusher to be crushed to obtain dendrobium officinale powder;
s22, adding deionized water into the reaction kettle, heating to 70 ℃, adding the dendrobium officinale powder obtained in the S21, controlling the rotating speed of the reaction kettle to be 600r/min, continuously stirring for 50min to obtain a mixed solution, filtering the mixed solution to remove residual dendrobium officinale particles, adding a compound enzyme into the mixed solution for enzymolysis, and controlling the enzymolysis time to be 5 h;
s23, carrying out adsorption and elution operations on the mixed solution after enzymolysis to obtain a semi-finished product solution, and finally concentrating the semi-finished product solution, wherein the concentration of the semi-finished product solution after concentration is controlled to be 35Brix, so that the dendrobium officinale extracting solution is obtained.
In this example, in S23, the mixed solution is preferably subjected to an adsorption operation using a molecular sieve, and the mixed solution is preferably subjected to an elution operation using an aqueous ethanol solution.
In this embodiment, it is preferable that the sterilization operation includes the following steps:
s31, injecting the primary essence liquid into a sterilized glass container, and sealing the glass container with a sealing cover;
s32, starting an ultraviolet sterilizer to sterilize the outside of the glass container and the sealing cover, opening the sealing cover, sending the glass container and the primary essence liquid into the sterilization equipment, and pumping out air in the sterilization equipment by using a vacuum pump to ensure that the inside of the sterilization equipment is in a vacuum state;
s33, starting the sterilization equipment to sterilize the primary essence liquid in the glass container, and simultaneously mixing the primary essence liquid by using the stirring mechanism in the sterilization equipment.
In this embodiment, it is preferable that in S33, the stirring speed of the stirring mechanism is controlled to 80r/min, and then the sterilization time of the sterilization apparatus is controlled to 5 h.
In this embodiment, the vitamin complex is preferably a combination of vitamin a, vitamin B6, vitamin E, and vitamin H, wherein the mass ratio of vitamin a, vitamin B6, vitamin E, and vitamin H is 1: 1: 2: 1.5.
in this embodiment, preferably, the plant polysaccharide is a combination of flammulina velutipes polysaccharide, tremella polysaccharide, acronychia negra polysaccharide, ginseng polysaccharide and cistanche polysaccharide, and the mass ratio of the flammulina velutipes polysaccharide to the tremella fuciformis polysaccharide to the ginseng polysaccharide to the cistanche polysaccharide is 2: 3: 2: 1: 2.
in this embodiment, preferably, the stem cell exosome suspension is a mesenchymal stem cell exosome suspension, and the stem cell exosome suspension contains a skin-improving plasmid vector.
In this embodiment, preferably, the stem cell exosome suspension is prepared as follows:
s41, culturing the stem cells by using a serum-containing culture medium, extracting after the culture is finished to obtain the stem cells, and then placing the stem cells in the culture medium for 3 days;
s42, filtering to remove stem cells in the culture medium, filtering, putting the filtered stem cell solution into a centrifuge tube, centrifuging, removing supernatant in the centrifuge tube to obtain a stem cell suspension, and then precipitating;
s43, inoculating the precipitated stem cell suspension into a culture bottle at the cell density of 50000 cells/CM 2, and then culturing the culture bottle;
s44, after the culture is finished, collecting the stem cell suspension in all culture bottles into a centrifugal container, repeatedly centrifuging for 5 times, removing supernatant after each centrifugation, and finally obtaining a precipitation solution, namely the stem cell exosome suspension.
Experiment:
preparing the essences prepared in examples 1-3 and conventional essences in the market, dividing each essence into 50 parts in equal amount, randomly inviting 200 test persons with the age of 40-60 years, distributing the essences prepared in examples 1, 2 and 3 and the conventional essences by taking 50 test persons as a group, establishing a WeChat group to record the use feedback of the test persons in real time, and calling the groups of the test persons in examples 1, 2, 3 and the conventional essences as 1 group, 2 group, 3 group and conventional group respectively;
the trial period is six months, the produced essence is averagely used twice per person every day, the using method is that the essence is evenly smeared on the skin after the face is cleaned, and finally, the statistical trial data is as follows:
group 1: 2 subjects quit midway among 50 subjects, and the satisfaction rate is 99.5 percent; the test person shows that the skin is not stimulated after long-term use, and the skin feels comfortable and is not greasy after use, so that the anti-wrinkle anti-aging skin care cream has better effects of resisting wrinkles, preventing aging, improving skin quality and whitening and protecting skin;
and 2, group: 5 quits from 50 test persons midway, and the satisfaction rate is 97.7%; the test person shows that the skin is not stimulated after long-term use, and the skin feels comfortable and is not greasy after use, so that the anti-wrinkle anti-aging skin care cream has better effects of resisting wrinkles, preventing aging, improving skin quality and whitening and protecting skin;
and 3, group: 3 test subjects quit midway among 50 test subjects, and the satisfaction rate is 98.6%; the test person shows that the skin is not stimulated after long-term use, and the skin feels comfortable and is not greasy after use, so that the anti-wrinkle anti-aging skin care cream has better effects of resisting wrinkles, preventing aging, improving skin quality and whitening and protecting skin;
conventional group: 18 test subjects quit midway through the method, and the satisfaction rate is only 68.3%; after the test, 15 test persons have symptoms of skin allergy, pruritus and the like after one month, and after the test for 6 months, the skin quality of the skin is not obviously improved.
The essence prepared in the embodiment 1, the embodiment 2 and the embodiment 3 has no side effect, no dependence and obvious anti-aging effect, and also has the effect of improving the skin; the conventional essence is not suitable for all skins, can generate symptoms such as allergy when used by some people, and has poor anti-aging effect.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. A preparation method of stem cell exosome anti-aging essence is characterized by comprising the following steps: the method comprises the following steps:
s11, preparing materials: preparing the following raw materials in parts by weight: 2-10 parts of stem cell exosome suspension, 5-10 parts of dendrobium officinale extract, 2-5 parts of ganoderma lucidum fermentation filtrate, 1-3 parts of vitamin complex, 0.5-0.8 part of schizophyllan, 1-3 parts of towel gourd extract, 1-3 parts of plant polysaccharide and 20-65 parts of deionized water:
s12, primary mixing: sequentially placing the dendrobium officinale extracting solution, the lucid ganoderma fermentation filtrate, the towel gourd extracting solution, the plant polysaccharide and the deionized water into a reaction kettle, controlling the rotating speed of the reaction kettle at 200-500r/min and the temperature at 40-65 ℃, and continuously mixing for 20-40min to obtain a mixed solution;
s13, secondary mixing: sequentially adding stem cell exosome suspension, multivitamins and schizophyllan into the mixed solution obtained in S12, controlling the temperature of the reaction kettle at 30-40 ℃, continuously mixing for 1-4h, and taking out to obtain the essence;
s14, sterilizing and filling: and (5) sterilizing the essence obtained in the step (S13), and filling the sterilized essence by using filling equipment, wherein the net content of each bottle is controlled to be 50-100 ml.
2. The method for preparing the anti-aging essence of stem cell exosomes according to claim 1, which is characterized in that: the suspension of the stem cell exosomes contains 20-35mg of protein per ml of solution.
3. The method for preparing the anti-aging essence of stem cell exosomes according to claim 1, which is characterized in that: the preparation method of the dendrobium officinale extracting solution comprises the following steps:
s21, selecting fresh and clean dendrobium officinale to dry, and then putting the dried dendrobium officinale into a crusher to be crushed to obtain dendrobium officinale powder;
s22, adding deionized water into the reaction kettle, heating to 50-70 ℃, adding the dendrobium officinale powder obtained in the S21, controlling the rotating speed of the reaction kettle to be 600r/min, continuously stirring for 20-50min to obtain a mixed solution, filtering the mixed solution to remove residual dendrobium officinale particles, adding a complex enzyme into the mixed solution for enzymolysis, and controlling the enzymolysis time to be 1-5 h;
s23, carrying out adsorption and elution operations on the mixed solution after enzymolysis to obtain a semi-finished product solution, and finally concentrating the semi-finished product solution to control the concentration of the semi-finished product solution after concentration to be 18-35Brix, thus obtaining the dendrobium officinale extracting solution.
4. The method for preparing the anti-aging essence of stem cell exosomes according to claim 3, wherein the anti-aging essence comprises the following steps: in the step S23, the mixed solution is adsorbed by using a molecular sieve, and the mixed solution is eluted by using an ethanol aqueous solution.
5. The method for preparing the anti-aging essence of stem cell exosomes according to claim 4, wherein the anti-aging essence comprises the following steps: the sterilization operation comprises the following specific steps:
s31, injecting the primary essence liquid into a sterilized glass container, and sealing the glass container with a sealing cover;
s32, starting an ultraviolet sterilizer to sterilize the outside of the glass container and the sealing cover, opening the sealing cover, sending the glass container and the primary essence liquid into the sterilization equipment, and pumping out air in the sterilization equipment by using a vacuum pump to ensure that the inside of the sterilization equipment is in a vacuum state;
s33, starting the sterilization equipment to sterilize the primary essence liquid in the glass container, and simultaneously mixing the primary essence liquid by using the stirring mechanism in the sterilization equipment.
6. The method for preparing the anti-aging essence of stem cell exosomes according to claim 1, which is characterized in that: and in the step S33, controlling the stirring speed of the stirring mechanism to be 50-80r/min, and then controlling the sterilization time of the sterilization equipment to be 3-5 h.
7. The method for preparing the anti-aging essence of stem cell exosomes according to claim 1, which is characterized in that: the compound vitamin is formed by combining vitamin A, vitamin B6, vitamin E and vitamin H, wherein the mass ratio of the vitamin A to the vitamin B6 to the vitamin E to the vitamin H is 1: 1: 2: 1.5.
8. the method for preparing the anti-aging essence of stem cell exosomes according to claim 1, which is characterized in that: the plant polysaccharide is formed by combining needle mushroom polysaccharide, tremella polysaccharide, emblic leafflower fruit polysaccharide, ginseng polysaccharide and desertliving cistanche polysaccharide, wherein the mass ratio of the needle mushroom polysaccharide to the tremella polysaccharide to the emblic leafflower fruit polysaccharide to the ginseng polysaccharide to the desertliving cistanche polysaccharide is 2: 3: 2: 1: 2.
9. the method for preparing the anti-aging essence of stem cell exosomes according to claim 1, which is characterized in that: the stem cell exosome suspension is a mesenchymal stem cell exosome suspension, and the stem cell exosome suspension contains a plasmid vector for improving skin.
10. The method for preparing the anti-aging essence of stem cell exosomes according to claim 1, which is characterized in that: the preparation method of the stem cell exosome suspension comprises the following steps:
s41, culturing the stem cells by using a serum-containing culture medium, extracting after the culture is finished to obtain the stem cells, and then placing the stem cells in the culture medium for culturing for 1-3 days;
s42, filtering to remove stem cells in the culture medium, filtering, putting the filtered stem cell solution into a centrifuge tube, centrifuging, removing supernatant in the centrifuge tube to obtain a stem cell suspension, and then precipitating;
s43, inoculating the precipitated stem cell suspension into a culture bottle at the cell density of 50000 cells/CM 2, and then culturing the culture bottle;
s44, after the culture is finished, collecting the stem cell suspension in all culture bottles into a centrifugal container, repeatedly centrifuging for 3-5 times, removing supernatant after each centrifugation, and finally obtaining a precipitation solution, namely the stem cell exosome suspension.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114344360A (en) * | 2022-01-07 | 2022-04-15 | 阿谷巴(杭州)生物科技发展有限公司 | Exosome compound preparation for skin photodamage repair and preparation method thereof |
| CN115645352A (en) * | 2022-11-21 | 2023-01-31 | 广州市科能化妆品科研有限公司 | Skin soothing and repairing composition and application thereof and repairing cream |
| CN117243885A (en) * | 2023-11-15 | 2023-12-19 | 北京青藤谷禧干细胞科技研究院有限公司 | Stem cell exosome composition for improving skin and preparation method thereof |
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| CN117243885B (en) * | 2023-11-15 | 2024-01-26 | 北京青藤谷禧干细胞科技研究院有限公司 | Stem cell exosome composition for improving skin and preparation method thereof |
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