CN115737523B - Female antibacterial repair care solution based on cell composite factors - Google Patents
Female antibacterial repair care solution based on cell composite factors Download PDFInfo
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Abstract
The invention discloses a female antibacterial repair care solution based on cell composite factors, which comprises the following components: stem cell complex factor, immune cell complex factor and water, wherein the concentration of the stem cell complex factor is 1 mu g/mL-5 mu g/mL, the concentration of the immune cell composite factor is 10 ng/mL-100 ng/mL, and the female antibacterial repair care solution based on the cell composite factor does not contain Chinese herbal medicine extract and disinfectant. According to the female antibacterial repair care solution based on the cell extract, traditional bactericides such as silver ions, chlorhexidine acetate and matrine are not added, stem cell complex factors and immune cell complex factors are selected, and the female antibacterial repair care solution can be used for a long time without long-term accumulated harmful effects by combining the results of a test example, and can achieve three effects of antibacterial, self-cleaning and repair.
Description
Technical Field
The invention relates to the technical field of nursing products, in particular to a female antibacterial repair nursing liquid based on cell composite factors.
Background
Investigation of the World Health Organization (WHO) into the health status of women worldwide has shown that: the global gynecological disease incidence rate is up to more than 65%, and in women of childbearing age, the gynecological disease incidence rate is over 70%; the number of attacks of genitourinary infections worldwide has reached 3.33 hundred million people each year. The general prevalence of common diseases in chinese women has increased slightly over the last decade. The total prevalence of common diseases in women nationwide in 2010 is 28.8%. Among the common diseases of women, colpitis is the first place.
Along with the rising of female population, the rising of working and living pressure, the rising of living standard and health consciousness and the like, gynecological diseases are more and more valued. Most of the time is in offices, on the way of going to and from work and on business, more time is in public places, and silk stockings are often worn, so that the air permeability is poor, and the probability of being infected by the outside is continuously increased. In addition, the working pressure is high, and the self resistance is reduced, so that nursing at private parts is more needed to be concerned at any time, and pathogen infection is avoided. For this purpose, a feminine care solution is typically used for cleaning care.
The prior private care liquid products are mainly divided into two types: the medicine is compounded by Chinese herbal medicine extract liquid such as matrine, phellodendron bark and the like, has the main function of treating gynecological diseases and plays a role of medicinal use; the other type is a disinfectant with sterilizing function, which mainly comprises silver ions, chlorhexidine acetate and other components with sterilizing function. Both products cannot be used for a long time from the aspect of components, and although the products have cleaning and sterilizing effects, the components have irritation to relatively fragile skin, and can kill too many normal bacterial groups, destroy the acidic environment, reduce the immunity of the organism and destroy the self-cleaning effect of the organism.
Disclosure of Invention
Based on the above, it is necessary to provide a female bacteriostatic repair care solution based on cell complex factors, which can solve the above problems.
A female antibacterial repair care solution based on cell composite factors comprises the following components: stem cell complex factor, immune cell complex factor and water, wherein the concentration of the stem cell complex factor is 1 mu g/mL-5 mu g/mL, the concentration of the immune cell composite factor is 10 ng/mL-100 ng/mL, and the female antibacterial repair care solution based on the cell composite factor does not contain Chinese herbal medicine extract and disinfectant.
In one embodiment, the stem cell complex factor is extracted after the umbilical cord mesenchymal stem cells are disrupted.
In one embodiment, the immunocytokine complex is extracted by disrupting umbilical cord blood mononuclear cells.
In one embodiment, the method further comprises the step of extracting the chlorella pumilus, wherein the concentration of the chlorella pumilus extract is 5-15 mug/mL.
In one embodiment, the chlorella pumila extract is a lecithin-coated chlorella pumila extract.
In one embodiment, the method further comprises chlorella extract, wherein the concentration of the chlorella extract is 5 mug/mL-15 mug/mL.
In one embodiment, the method further comprises the step of extracting the rhodococcus rhodochrous, wherein the concentration of the rhodochrous extract is 2-10 mug/mL.
In one embodiment, the method further comprises fructooligosaccharides with a concentration of 100-300 μg/mL and beta-glucan with a concentration of 3-20 μg/mL.
In one embodiment, the water treatment agent further comprises parahydroxybenzene acrylic acid, hyaluronic acid and seawater, wherein the concentration of the parahydroxybenzene acrylic acid is 20-100 mug/mL, and the volume ratio of the seawater to the water is 1-5: 99 to 95 percent.
In one embodiment, the herbal extract comprises matrine extract and phellodendron extract, and the disinfectant comprises silver ions and chlorhexidine acetate.
According to the female antibacterial repair care solution based on the cell extract, traditional bactericides such as silver ions, chlorhexidine acetate and matrine are not added, stem cell complex factors and immune cell complex factors are selected, and the female antibacterial repair care solution can be used for a long time without long-term accumulated harmful effects by combining the results of a test example, and can achieve three effects of antibacterial, self-cleaning and repair.
Furthermore, the female antibacterial repair care solution based on the cell extract also comprises the chlorella pumila extract, the chlorella vulgaris extract and the chlorella vulgaris extract, wherein the algae extract can regulate the prebiotics of vaginal flora, relieve allergy and resist inflammation, and the algae extract, the stem cell complex factor and the immune cell complex factor are combined, so that the female antibacterial repair care solution can be used for a long time, can not produce long-term accumulated harmful effects, and can achieve three effects of bacteriostasis, self-cleaning and repair.
It should be noted that a large number of microorganisms including bacteria, fungi, viruses, etc. exist in the private parts of women, and these microorganisms play an important role in maintaining the ecological balance of skin, self-purifying effect and participating in physiological functions of skin mucosa, and when local immunity is lowered or the private parts are infected by external microorganisms, the microorganisms become dysbiosis, and they may become pathogenic bacteria to cause diseases, and the accumulation and non-metabolizability caused by the disorder of conventional bactericides and antibiotics also cause damage to beneficial bacteria, further dysbacteriosis, and possible recurrent inflammation attack. Restoring and maintaining normal microbial colony, improving local immunity of private parts and repairing small damage of mucous membrane at private parts in time has important significance for keeping health of private parts and preventing external bacteria infection.
According to the results of the test examples, the female antibacterial repair care solution based on the cell extract has the advantages of good cleaning antibacterial effect, mild and non-irritating skin at private parts of females, nutrition and repair effects, and convenience in daily use.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The invention discloses a female antibacterial repair care solution based on cell composite factors, which comprises the following components: the concentration of the stem cell composite factor is 1 mu g/mL-5 mu g/mL, the concentration of the immune cell composite factor is 10 ng/mL-100 ng/mL, and the female antibacterial repair care solution based on the cell composite factor does not contain Chinese herbal medicine extract and disinfectant.
In the present invention, the concentration of the stem cell complex factor refers to the total protein concentration of the stem cell complex factor, and the concentration of the immune cell complex factor refers to the protein concentration of the immune cell complex factor.
According to the female antibacterial repair care solution based on the cell extract, traditional bactericides such as silver ions, chlorhexidine acetate and matrine are not added, stem cell complex factors and immune cell complex factors are selected, and the female antibacterial repair care solution can be used for a long time without long-term accumulated harmful effects by combining the results of a test example, and can achieve three effects of antibacterial, self-cleaning and repair.
Specifically, the Chinese herbal medicine extract comprises matrine extract and phellodendron extract, and the disinfectant comprises silver ions and chlorhexidine acetate.
Namely, the female antibacterial repair care solution based on the cell extract does not contain matrine extract, phellodendron extract, silver ions and chlorhexidine acetate.
In addition, it is pointed out that the female antibacterial repair care solution based on the cell extract does not contain other types of Chinese herbal medicine extract and disinfectant.
Preferably, in the present invention, the stem cell complex factor is extracted by crushing umbilical mesenchymal stem cells.
The cell composite factors extracted after umbilical cord mesenchymal stem cells are crushed, including Epidermal Growth Factor (EGF), stem Cell Factor (SCF), basic fibroblast growth factor (bFGF), keratinocyte Growth Factor (KGF), vascular Endothelial Growth Factor (VEGF), insulin-like growth factor (IGF), transforming growth factor-beta 1 (TGF-beta 1) and the like, can promote the self repair of injured tissues and cells, regulate local immune response and inhibit inflammatory response.
Preferably, in the present invention, the immunocytokine complex is obtained by extracting umbilical cord blood mononuclear cells after disruption.
The immunocyte complex factor extracted from the crushed umbilical cord blood mononuclear cells can strengthen local immunity and resist invasion of external bacteria.
Preferably, the female antibacterial repair care solution based on the cell extract further comprises a chlorella pumilus extract, wherein the concentration of the chlorella pumilus extract is 5-15 mug/mL.
Specifically, the chlorella pumila extract is a lecithin-coated chlorella pumila extract.
The chlorella pumila extract obtained by rescreening from a plurality of plankton is wrapped by fat-soluble lecithin, and the wrapped microcapsule can go deep into skin to relieve allergy and resist inflammation. By reducing the amount of pro-inflammatory interleukins that cause abnormal reactions in leukocytes, the skin is calmed and relaxed, and redness, swelling and itching are reduced. By enhancing the defense system of the skin, the secretion of ceramide is increased, the skin barrier is enhanced, the immune system is enhanced to resist external invasion, and the skin which is easy to be allergic is helped. By modulating the reduction of the pro-inflammatory interleukins IL-1 beta and IL-8, calm and sooth the skin. Through regulating IFN-gamma (IFN-gamma), the protection system of people is enhanced, the secretion of ceramide is increased, the barrier of skin is finally enhanced, the barrier of skin is enhanced, the immune system is enhanced against external invasion, and the skin which is easy to be allergic is helped. By modulating the reduction of IL2 (IL 2 has an anti-itching effect, which is intimately associated with allergic inflammation, severe skin itching and skin dryness), the symptoms of itching, rash and redness are reduced.
Preferably, the female antibacterial repair care solution based on the cell extract further comprises chlorella extract, wherein the concentration of the chlorella extract is 5-15 mug/mL.
The chlorella contains a high-efficiency repairing factor, has high activity and can provide more nutrition for skin. The plant-derived repair factor can restore cell skin activity, help renew keratinocytes, promote deep cell growth, and restore damaged skin surface barrier. Upregulation of the endogenous growth factor TGF-alpha and the skin thickening signal factor DKK-1.
Preferably, the female antibacterial repair care solution based on the cell extract further comprises a chlorella extract, wherein the concentration of the chlorella extract is 2-10 mug/mL.
The extract of Synechococcus which is extracted from natural Synechococcus can significantly increase the moisture content of skin, and can be kept for 24 hours. And can improve skin texture, relieve roughness, and fine and smooth skin.
The female antibacterial repair care solution based on the cell extract also comprises the chlorella pumilus extract, the chlorella vulgaris extract and the chlorella vulgaris extract, wherein the algae extract can regulate the prebiotics of vaginal flora, relieve allergy and resist inflammation, and the combination of the algae extract, the stem cell complex factor and the immune cell complex factor can be used for a long time, does not produce long-term accumulated harmful effects, and can achieve three effects of bacteriostasis, self-cleaning and repair.
Preferably, the female antibacterial repair care solution based on the cell extract further comprises fructo-oligosaccharide and beta-glucan, wherein the concentration of the fructo-oligosaccharide is 100-300 mug/mL, and the concentration of the beta-glucan is 3-20 mug/mL.
The fructo-oligosaccharide can regulate the surface flora of skin, promote the activity of beneficial bacteria on the body surface, inhibit harmful bacteria and restore the microecological environment on the body surface. Providing nutrition for skin and epithelial cells, maintaining cell viability, promoting repair, recovering skin barrier function, increasing immunity, and ensuring skin to adapt to environment and life style.
Preferably, the female antibacterial repair care solution based on the cell extract further comprises p-hydroxy-phenyl-acrylic acid, hyaluronic acid and seawater, wherein the concentration of the p-hydroxy-phenyl-acrylic acid is 20-100 mug/mL, and the volume ratio of the seawater to the water is 1-5: 99 to 95 percent.
The active ingredient p-hydroxy-phenyl-acrylamide-benzoic acid extracted from oat can obviously relieve abnormal conditions of skin, relieve inflammatory response caused by histamine release, and rapidly relieve itching, redness and blisters caused by various conditions.
It should be noted that a large number of microorganisms including bacteria, fungi, viruses, etc. exist in the private parts of women, and these microorganisms play an important role in maintaining the ecological balance of skin, self-purifying effect and participating in physiological functions of skin mucosa, and when local immunity is lowered or the private parts are infected by external microorganisms, the microorganisms become dysbiosis, and they may become pathogenic bacteria to cause diseases, and the accumulation and non-metabolizability caused by the disorder of conventional bactericides and antibiotics also cause damage to beneficial bacteria, further dysbacteriosis, and possible recurrent inflammation attack. Restoring and maintaining normal microbial colony, improving local immunity of private parts and repairing small damage of mucous membrane at private parts in time has important significance for keeping health of private parts and preventing external bacteria infection.
According to the results of the test examples, the female antibacterial repair care solution based on the cell extract has the advantages of good cleaning antibacterial effect, mild and non-irritating skin at private parts of females, nutrition and repair effects, and convenience in daily use.
The following are specific examples.
In a specific embodiment, the drug sources involved are as follows:
the Chlorella extract is available from Spain green (L.) Inc BioSKN；
The Chlorella extract is HYDRINTENSE of GivaudanFranceSAS in Switzerland;
The Chlorella pumilus extract was purchased from Spanish green hopper.L. and the fructooligosaccharides were purchased from Shandong mountain Biotechnology Co., ltd
Beta-glucan was purchased from Beijing Sanyou Hui Biotechnology Co., ltd
Para-hydroxy-phenylacrylaminobenzoic acid was purchased from De Xin (Shanghai) Inc
Example 1: extraction of umbilical cord mesenchymal stem cell composite factor
Cell resuscitation
1) 1 Human umbilical cord mesenchymal stem cell (2× 7) extracted from umbilical cord mesenchymal stem cells public purse of a healthy donor is taken out, water-bath is carried out in a water bath kettle at 37 ℃, light shaking is carried out, and ice blocks are taken out when ice blocks in a freezing tube are about to be completely melted. And (5) rapidly carrying out surface disinfection on the freezing tube and then transferring the tube to a biological safety cabinet. The resuscitated cell suspension was aspirated and transferred into 40mL of physiological saline which had been pre-chilled at 4 ℃.500g centrifugal force, 10 minutes, 9 speed up, 7 speed down, centrifugal force.
2) After centrifugation, the mixture was washed again with 40mL of physiological saline, and 20mL of the mixture was transferred to another separation tube. 500g,10 min, 9 rise, 7 fall, and centrifugation.
3) And (5) centrifuging for later use.
Ultrasonic crushing
1) The prepared cells were resuspended in 15mL of physiological saline, transferred to a 50mL centrifuge tube, and the tube placed in ice.
2) HN-250M hand-held ultrasonic cell grinder settings: horn gauge: phi 3; power: 25%, ultrasonic time: 3s, ultrasonic gap time: 2s, total ultrasound time: 2h.
3) After completion of the crushing, 500. Mu.L was centrifuged at 5000rpm for 10 min.
4) After centrifugation was completed, the supernatant was retained.
5) The total protein concentration in the supernatant was measured to be 13.3. Mu.g/mL using a Nanodrop2000 spectrophotometer.
Protein concentration
The supernatant was transferred to a 50mL ultrafiltration centrifuge tube (Millipore), centrifuged at 4000g for 45min, at 9 speed, at 7 speed, and centrifuged. And after the centrifugation is finished, obtaining the umbilical cord mesenchymal stem cell composite factor solution.
The total protein concentration of the umbilical cord mesenchymal stem cell composite factor solution was 167.8 mug/mL as measured by using a Nanodrop2000 spectrophotometer.
Example 2: extraction of umbilical cord blood mononuclear cell composite factor
1) Centrifuging 80mL of neonatal umbilical cord blood of a healthy donor for 8min under the centrifugal force of 500g, sucking an upper pale yellow plasma layer by a pipette after centrifugation, transferring the upper pale yellow plasma layer into a new 50mL centrifuge tube, and inactivating the obtained plasma for 30min at 56 ℃ to obtain inactivated umbilical cord plasma for later use;
2) The rest liquid is concentrated blood cells, the concentrated blood cells are diluted by normal saline, the concentrated blood cells are centrifugally separated by density gradient of human lymphocyte separation liquid, the centrifugation is carried out for 20min under 840g centrifugal force, the lower layer of white membrane layer (UCBMC, umbilical cord blood mononuclear cells) is sucked, and after the resuspension by the normal saline, the centrifugation is carried out for two times by 300g centrifugal force and 8min to obtain UCBMC;
3) Inoculating UCBMC cells obtained into a lymphocyte serum-free medium, performing amplification culture on UCBMC cells by using the inactivated umbilical cord plasma obtained in 1) according to the volume concentration of 10% and IL-2 pair of 300U/mL at 37 ℃ and under the condition of 5% CO 2 for 72 hours, and finally performing centrifugal separation to collect cells and supernatant respectively;
4) Re-suspending the cells obtained in the step 3) into 1mL of physiological saline, transferring the physiological saline into 1mL of freezing pipes, placing the freezing pipes on a freezing pipe rack, placing the freezing pipe rack into a foam box, adding ice into the foam box, and paying attention that the height of the ice cannot exceed the height of the freezing pipes; placing the foam box into a biosafety cabinet after sterilizing, and removing the freezing storage tube cover;
5) An HN-250M handheld ultrasonic cell crusher is used in a biosafety cabinet, an amplitude transformer is inserted 10mm below the cell suspension liquid level of a freezing storage tube, and the amplitude transformer cannot contact the bottom or the tube wall of the freezing storage tube;
6) Ultrasonic breaker sets up: power: 30%, ultrasonic time: 3s, ultrasonic gap time: 2s, total ultrasound time: 4h.
7) After cell disruption, the cell debris and factor solution were aspirated with a pipette, transferred to a 1.5mL centrifuge tube, spun at 10000rpm for 10min, and centrifuged.
8) After centrifugation, the supernatant was aspirated, transferred to a 2mL cryopreservation tube, and physiological saline was added to 1mL to obtain cytokine solution 1. The total protein concentration in cytokine solution 1 was measured to be 26.4 μg/mL using a Nanodrop2000 spectrophotometer.
9) Transferring the supernatant obtained in the step 3) into an ultrafiltration centrifuge tube, 4000g,30min, upshift to 9 th gear, downshift to 7 th gear, centrifuge.
10 After centrifugation, sucking out the concentrated factor solution by using a 100 mu L gun head, transferring the concentrated factor solution into a 2mL freezing tube, adding 500 mu L physiological saline into an ultrafiltration centrifuge tube, washing a filter membrane component, sucking out the concentrated factor solution by using the 100 mu L gun head, transferring the concentrated factor solution into the same freezing tube, adding physiological saline into the same freezing tube to adjust the volume to 1mL, and obtaining the cytokine solution 2, wherein the total protein concentration in the cytokine solution 2 is 11.7 mu g/mL by using a Nanodrop2000 spectrophotometer.
11 0.08ML of cytokine solution 1 and 0.02mL of cytokine solution 2 are mixed to obtain the umbilical cord blood mononuclear cell composite factor solution.
Example 3
80ML of water was added to the main kettle, heated to 82.5.+ -. 2.5 ℃ with stirring, and stirred at constant temperature.
Cooling to 42.5deg.C+ -2.5deg.C under stirring, adding 1mL of fructooligosaccharide solution, 1mL of beta-glucan solution, 1mL of Chlorella extract solution, 1mL of Chlorella floating extract solution, 0.21mL of p-hydroxyphenylacrylamide benzoic acid, 1mL of hyaluronic acid, 5mL of seawater, 3mL of umbilical cord mesenchymal stem cell complex factor solution and 0.1mL of umbilical cord blood mononuclear cell complex factor solution, and stirring.
The cooling water was started while stirring and cooled to 32.5.+ -. 2.5 ℃.
And (5) filling after sampling and detecting to be qualified, and obtaining the female antibacterial repair care solution based on the cell extract.
Test case
10 Female subjects enrolled in women with colpitis aged 30-60 years were sprayed twice daily in the morning and evening with the cell extract-based female bacteriostatic repair solution of example 3 for one month, and the results are shown in table 1 below.
TABLE 1
As can be seen from table 1, the female antibacterial repair care solution based on the cell extract in example 3 can be used for a long time, does not have long-term accumulated harmful effects, and can achieve three effects of antibacterial, self-cleaning and repair.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the claims. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (2)
1. A female antibacterial repair care solution based on cell composite factors is characterized by comprising the following components: stem cell complex factor, immune cell complex factor and water, wherein the concentration of the stem cell complex factor is 1 mu g/mL-5 mu g/mL, the concentration of the immune cell composite factor is 10 ng/mL-100 ng/mL, and the female antibacterial repair care solution based on the cell composite factor does not contain a Chinese herbal medicine extract and a disinfectant;
The immune cell composite factor is prepared by the following operation:
1) Centrifuging 80mL of neonatal umbilical cord blood for 8min under the centrifugal force of 500g, sucking an upper pale yellow plasma layer by a pipette after centrifugation, transferring the upper pale yellow plasma layer into a new 50mL centrifuge tube, and inactivating the obtained plasma for 30min at 56 ℃ to obtain inactivated umbilical cord plasma for later use;
2) The rest liquid is concentrated blood cells, the concentrated blood cells are diluted by normal saline, the concentrated blood cells are centrifugally separated by density gradient of human lymphocyte separation liquid, the centrifugation is carried out for 20min under 840g centrifugal force, the white membrane layer at the lower layer is sucked, and after the resuspension by the normal saline, the centrifugation is carried out for 300g centrifugal force and 8min centrifugal washing for two times, thus obtaining the umbilical blood mononuclear cells;
3) Inoculating the obtained cord blood mononuclear cells into a lymphocyte serum-free medium, then carrying out amplification culture on the cord blood mononuclear cells for 72 hours at 37 ℃ under the condition of 5% CO 2 by using the inactivated cord blood plasma obtained in 1) according to the volume concentration of 10% and interleukin-2 of 300U/mL, and finally carrying out centrifugal separation to collect cells and supernatant respectively;
4) Re-suspending the cells obtained in the step 3) into 1mL of physiological saline, transferring the physiological saline into 1mL of freezing pipes, placing the freezing pipes on a freezing pipe rack, placing the freezing pipe rack into a foam box, adding ice into the foam box, and paying attention that the height of the ice cannot exceed the height of the freezing pipes; placing the foam box into a biosafety cabinet after sterilizing, and removing the freezing storage tube cover;
5) An HN-250M handheld ultrasonic cell crusher is used in a biosafety cabinet, an amplitude transformer is inserted 10mm below the cell suspension liquid level of a freezing storage tube, and the amplitude transformer cannot contact the bottom or the tube wall of the freezing storage tube;
6) Ultrasonic breaker sets up: power: 30%, ultrasonic time: 3s, ultrasonic gap time: 2s, total ultrasound time: 4h;
7) After the cell disruption, sucking out cell fragments and factor solution by using a pipetting gun, transferring the cell fragments and factor solution into a 1.5mL centrifuge tube, centrifuging at 10000rpm for 10 min;
8) After centrifugation, sucking out the supernatant, transferring to a 2mL cryopreservation tube, and adding physiological saline to 1mL to obtain cytokine solution 1;
9) Transferring the supernatant obtained in the step 3) into an ultrafiltration centrifuge tube, carrying out up-speed 9, down-speed 7 and centrifugation on 4000g for 30min;
10 After centrifugation, sucking out the concentrated factor solution by using a 100 mu L gun head, transferring the concentrated factor solution into a 2mL freezing tube, adding 500 mu L physiological saline into an ultrafiltration centrifuge tube, washing a filter membrane component, sucking out the concentrated factor solution by using the 100 mu L gun head, transferring the concentrated factor solution into the same freezing tube, and adding physiological saline into the same freezing tube to adjust the volume to 1mL to obtain a cytokine solution 2;
11 Mixing 0.08mL of cytokine solution 1 and 0.02mL of cytokine solution 2 to obtain immune cell composite factor;
the stem cell composite factor is prepared by the following operations:
Cell resuscitation
1) Taking out 1 human umbilical cord mesenchymal stem cells (2× 7) extracted through umbilical cords of healthy donors, carrying out water bath in a water bath kettle at 37 ℃ and gently shaking, taking out when ice cakes in a freezing tube are about to be completely melted, carrying out surface sterilization on the freezing tube, transferring the sterilized cell suspension to a biosafety cabinet, sucking out the resuscitated cell suspension, transferring the resuscitated cell suspension into 40mL of physiological saline precooled at 4 ℃, carrying out centrifugal force of 500g for 10 minutes, increasing the speed by 9, reducing the speed by 7, and centrifuging;
2) After centrifugation, washing with 40mL of physiological saline again, taking 20mL and transferring to another separation tube; 500g,10 minutes, 9 speed up, 7 speed down, and centrifugation;
3) Centrifuging for later use;
Ultrasonic crushing
1) The prepared cells are resuspended in 15mL of physiological saline, transferred to a 50mL centrifuge tube, and the centrifuge tube is placed in ice;
2) HN-250M hand-held ultrasonic cell grinder settings: horn gauge: phi 3; power: 25%, ultrasonic time: 3s, ultrasonic gap time: 2s, total ultrasound time: 2h;
3) After the completion of the crushing, 500. Mu.L was taken, centrifugation at 5000rpm for 10 min;
4) After centrifugation is completed, the supernatant is retained;
Protein concentration
Transferring the supernatant into a 50mL ultrafiltration centrifuge tube, and obtaining stem cell complex factors after centrifugation, wherein the centrifugal force is 4000g, the centrifugal force is 45min, the speed is 9, the speed is 7, and the centrifugation is completed;
the chlorella pumilus also comprises chlorella pumilus extract, wherein the concentration of the chlorella pumilus extract is 5-15 mug/mL;
the chlorella extract also comprises chlorella extract, wherein the concentration of the chlorella extract is 5-15 mug/mL;
the chlorella extract also comprises a chlorella extract, wherein the concentration of the chlorella extract is 2-10 mug/mL;
the fructo-oligosaccharide and beta-glucan are also included, the concentration of the fructo-oligosaccharide is 100-300 mug/mL, and the concentration of the beta-glucan is 3-20 mug/mL;
The water treatment agent also comprises parahydroxybenzene acrylic acid, hyaluronic acid and seawater, wherein the concentration of the parahydroxybenzene acrylic acid is 20-100 mug/mL, and the volume ratio of the seawater to the water is 1-5: 99-95;
the Chinese herbal medicine extracting solution comprises matrine extracting solution and phellodendron extracting solution, and the disinfectant comprises silver ions and chlorhexidine acetate.
2. The cell complex factor-based female bacteriostatic repair care solution according to claim 1, wherein the chlorella pumila extract is a lecithin-coated chlorella pumila extract.
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| CN114652748A (en) * | 2022-03-29 | 2022-06-24 | 湖南有美生物科技有限公司 | Preparation method and application of medical gynecological lotion containing stem cell bacteriostatic factor |
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| CN107625680A (en) * | 2017-09-26 | 2018-01-26 | 杭州梵琳科技有限公司 | Shu Min aquas and preparation method thereof |
| 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 |
| CN111214431A (en) * | 2020-02-28 | 2020-06-02 | 深圳市芙丽嘉生物科技有限公司 | Composition for promoting skin recovery and application thereof |
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