CN103385890A - A preparation method for an anti-aging biological cell preparation - Google Patents
A preparation method for an anti-aging biological cell preparation Download PDFInfo
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- CN103385890A CN103385890A CN2013102988783A CN201310298878A CN103385890A CN 103385890 A CN103385890 A CN 103385890A CN 2013102988783 A CN2013102988783 A CN 2013102988783A CN 201310298878 A CN201310298878 A CN 201310298878A CN 103385890 A CN103385890 A CN 103385890A
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Abstract
A preparation method for an anti-aging biological cell preparation. The peripheral blood of healthy adults is used as a raw material for the anti-aging biological cell preparation which is prepared by the following steps: a, acquisition of the peripheral blood of healthy adults, which is used as the raw material for the preparation; b, separation and extraction of mononuclear cells in the peripheral blood; c, transformation and proliferation of the mononuclear cells after induction by cell mitogen in vitro; and d, detection of the obtained preparation. By being injected in a patient in an inoculation method of subcutaneous multi-point injection, the preparation of the invention can stimulate a body's immune cells, regulating immune function, effectively eliminate a sub-health state, delay dysfunction and be anti-aging, clinically achieving curative effects of adjusting the menstrual cycle, improving diet and sleep quality, promoting vitality and physical recovery, improving mental status, increasing disease resistance, making the skin bright and shiny, and maintaining a young appearance.
Description
Technical field
The present invention relates to a kind of biological preparation, particularly, by to the animal economy conditioning, treat women's subhealth state, delay hypokinetic defying age biological cell preparation, belong to medical technical field.
Background technology
Be subjected to the impact of special physiological, psychology, environmental factors, the adult female particularly enters the women in middle age, miscarriage or childbearing history are arranged more, under the interference of life and work pressure, mental pressure, environmental pollution, the various physiological functions of body descend gradually, the ovarian function decline, hormonal readiness changes, and causes occurring subhealth state and hypofunction and the old and feeble signs such as the dark and gloomy tarnish of skin, menoxenia and dysmenorrhea, inappetence, listless, poor sleeping quality, neurasthenia, disease-resistant and recovery capability reduction.Now, the various technical methods that beauty culture adopts, its objective is make the women retain youth beautiful appearance, cover the vestige in years, sub-health state, hypofunction and aging are looks improving and the skin nourishing, the most basic, most important, the most direct beautiful unfavorable factor retains youth.No matter be daily basic aesthetic nursing, beauty apparatus treatment; or the cosmetic formulation that sheep placental extract fashionable for a time, hyaluronic acid, creotoxin A etc. are advanced; be all to start with from part, where go wrong and where repair, belong to the method for curing the symptoms, not the disease of " treat the head when the head aches, foot bitterly cure foot ".At present, still lack and a kind ofly from whole conditioning human body function, start with, to the medical preparation that causes women's subhealth state and old and feeble root to be intervened reverse.
Summary of the invention
Technical problem to be solved by this invention is: overcome the defect of above-mentioned prior art and provide a kind of by body being carried out integral body conditioning, reach women's shining preparation method of adopting the defying age biological cell preparation of effect of preserving youthful looks.
The alleged problem of the present invention is solved by following technical scheme:
A kind of preparation method of defying age biological cell preparation, its special feature is: described defying age biological cell preparation is take the peripheral blood of health adult as raw material, and is prepared in the steps below:
A. take the healthy adult human peripheral obtained as described preparation raw material;
B. above-mentioned peripheral blood raw material is carried out the mononuclearcell separation and Extraction;
The human peripheral blood single nucleus cell that c. will separate is proliferation after the former phytohaemagglutinin of cell division (PHA) stimulated in vitro;
D. preparation detects.
The preparation method of above-mentioned defying age biological cell preparation, in described b step, get peripheral blood 15mL-20mL, after using Hank ' s liquid or PBS buffer 1-1.5 doubly to dilute, adopts lymphocyte separation medium to separate, collect mononuclearcell through the density gradient centrifugation; In described c step, get the PERIPHERAL BLOOD MONONUCLEAR CELL that the b step is separated, after Hank ' s liquid or PBS buffer solution for cleaning 2 times, take the RPMI-1640 complete culture solution, adjust cell concentration as 2 * 10
6/ mL, add phytohaemagglutinin (PHA) to make its final concentration to 100 μ g/mL, mixes rearmounted 37 ℃, 5%CO
2Cultivate altogether 72h in saturated humidity, after the centrifugal 5min of 1500rpm, abandon supernatant and obtain sedimentation cell,, adjustment cell concentration resuspended take the injection normal saline is 0.5 ~ 1 * 10
7/ mL, namely make described preparation.
The preparation method of above-mentioned defying age biological cell preparation, in described preparation detecting step,, with the preparation that the c step is made,, with the centrifugal 5min of 1500rpm, abandon supernatant and obtain sedimentation cell; Carry out viable count, trypanblue exclusion method judgement cell viability, cell smear HE staining oil Microscopic observation lymhocyte transformation rate, mtt assay under inverted microscope and detect cell activation propagation
A 492Value.Wherein, living cells ratio 〉=95%, lymhocyte transformation rate 〉=70%,
A 492Value between 0.85～1.20, cellular immune function is normal.
Defying age biological cell preparation of the present invention be peripheral blood take health adult as raw material, through the separation and Extraction of PERIPHERAL BLOOD MONONUCLEAR CELL, the steps such as external evoked proliferation of isolated cell, be prepared from, described preparation is a kind of cytobiology preparation of transferring body's immunity, the women being called forth one's youthful vigor again of nursing one's health by integral body.The vaccination ways of preparation of the present invention with subcutaneous multi-point injection is expelled in patient body, can be by stimulating the immunocyte of body, regulate immunologic function, effectively treat sub-health state, delay hypofunction and defying age, reach clinically the curative effect of adjusting menstrual cycle, improving diet sleep quality, promotion vigor and antisecosis, improve mental status, improve resistance against diseases, make the beautiful gloss of skin, preserve youthful looks and shine and adopt.Preparation of the present invention is pure cell component,, without complementary substance, uses safe and effectively, and curative effect is sure, and clinical observation has no short-term and long-term untoward reaction and toxic and side effects.
The specific embodiment
The invention provides a kind of take the preparation method of health adult's peripheral blood as raw-material defying age biological cell preparation.Described preparation is by body being carried out integral body conditioning, treatment women sub-health state, delays hypofunction and defying age, thereby makes the beautiful gloss of female skin, shining the adopting of preserving youthful looks.Research is found, after defying age biological cell preparation subcutaneous injection provided by the invention, the lymphocytic quantity of body T, activation signals transduction level, activation and proliferation ability are significantly improved, promote forward immunological effect and negative regulation T lymphocyte balance, the collaborative stimulating factor of positive negative sense to express balance, Th1/ Th2 cytokine-expressing balance, thus stimulate body immunocyte, regulate whole immunologic function.
The preparation method of defying age biological cell preparation of the present invention is as follows:
1, get the peripheral blood 15mL-20mL of health adult that meets the blood donation personnel national standard, with Hank ' s liquid or PBS buffer 1-1.5, doubly dilute; With lymphocyte separation medium through density gradient centrifugation separated and collected mononuclearcell.
The mononuclearcell that 2, will separate is through the former external Induction Transformation propagation of cell division: get the mononuclearcell of above-mentioned separated and collected, after Hank ' s liquid or PBS buffer solution for cleaning 2 times, take the RPMI-1640 complete culture solution, adjust cell concentration as 2 * 10
6/ mL, the mitogen that uses is phytohaemagglutinin (Phytohemagglutinin, PHA), stimulates final concentration to 100 μ g/mL; The condition of hatching altogether with cell is for mixing rearmounted 37 ℃, 5%(volume unit) CO
2Cultivate 72h in saturated humidity, after the centrifugal 5min of 1500rpm, abandon supernatant and obtain sedimentation cell,, adjustment cell concentration resuspended take the injection normal saline is 0.5 ~ 1 * 10
7/ mL, namely make described preparation.
3, the detection of described preparation is as follows: above-mentioned preparation, with the centrifugal 5min of 1500rpm, is abandoned supernatant and obtained sedimentation cell; Carry out viable count, trypanblue exclusion method judgement cell viability, cell smear HE staining oil Microscopic observation lymhocyte transformation rate, mtt assay under inverted microscope and detect cell activation propagation
A 492Value; Wherein, living cells ratio 〉=95%, the lymphocyte transformation more than 70% are lymphoblast, occur that volume increases, endochylema is abundant, basophilia strengthens, nuclear membrane is clear and the loose morphological change that is the typical proliferation such as netted, visible kernel of pseudopodium sample projection, chromatin is arranged;
A 492Value between 0.85 ~ 1.20, cellular immune function is normal, mtt assay detects cell activation propagation
A 492Value raise and to have extremely significant significant difference (compare with the cellular control unit of without the PHA activation, inducing,
P＜0.0005).Described cellular immune function mainly comprises lymhocyte transformation rate, Th
1/ Th
2Cytokine secretion and cell surface CD3
+, CD4
+, CD8
+, IL-2R α
+, CD4
+CD25
+, CD4
+CD28
+, CD4
+CTLA-4
+Positive expression rate etc.
When preparation of the present invention used, described preparation is resuspended with the injection normal saline, and adjusting cell concentration was 0.5 ~ 1 * 10
7/ mL; Get 2mL and treat with the vaccination ways of subcutaneous multi-point injection, in every minor tick 3-4 week, 3 times is a course for the treatment of, and curative effect can be kept about 0.5-1.
, for guaranteeing safety and the effectiveness of preparation of the present invention, require the whole blood source to be normal adults, to meet the blood donation personnel national standard; Preparation prepares omnidistance sterile working, and the vessel articles for use that use, buffer, separating medium, cleanout fluid, injection liquid meet immunology operation and the aseptic requirement of pharmaceuticals industry.
Preparation curative effect of the present invention is pressed following Standard Judgement: with subhealth state and old and feeble sign, improve as treatment effectively.Namely by the dark and gloomy tarnish of skin, menoxenia and the dysmenorrhea that occurred, inappetence, listless, poor sleeping quality, neurasthenia, disease-resistant and recovery capability reduction etc. in the past, through treatment change the beautiful gloss of skin, menstrual cycle rule into and dysmenorrhea weakens or disappearance, diet sleep quality obviously improve, mental status is good and energetic, antisecosis is fast, recovery capability obviously improves etc. after disease-resistant and disease.Table 1, for after clinical, improves sub-health state, senile-resistant efficacy statistical table for preparation of the present invention.
Table 1
Claims (3)
1. the preparation method of a defying age biological cell preparation is characterized in that: described defying age biological cell preparation is take the peripheral blood of health adult as raw material, and its preparation method carries out in the steps below:
A. extracting the healthy adult human peripheral is described preparation raw material;
B. above-mentioned peripheral blood raw material is carried out the mononuclearcell separation and Extraction;
The human peripheral blood single nucleus cell that c. will separate is proliferation after the former phytohaemagglutinin of cell division (PHA) stimulated in vitro;
D. preparation detects.
2. the preparation method of defying age biological cell preparation according to claim 1, it is characterized in that: in described b step, get peripheral blood 15mL-20mL, use Hank ' s liquid or PBS buffer 1-1.5 doubly to dilute, adopt lymphocyte separation medium through density gradient centrifugation separated and collected mononuclearcell; In described c step, get the PERIPHERAL BLOOD MONONUCLEAR CELL that the b step is separated, after Hank ' s liquid or PBS buffer solution for cleaning 2 times, take the RPMI-1640 complete culture solution, adjust cell concentration as 2 * 10
6/ mL, add PHA-P HA to make its final concentration to 100 μ g/mL, mixes rearmounted 37 ℃, 5%CO
2Cultivate altogether 72h in saturated humidity, after the centrifugal 5min of 1500rpm, abandon supernatant and obtain sedimentation cell,, adjustment cell concentration resuspended take the injection normal saline is 0.5 ~ 1 * 10
7/ mL, namely make described preparation.
3. the preparation method of defying age biological cell preparation according to claim 2, is characterized in that: in described preparation detecting step,, with the preparation of making in the c step,, with the centrifugal 5min of 1500rpm, abandon supernatant and obtain sedimentation cell; Carry out viable count, trypanblue exclusion method judgement cell viability, cell smear HE staining oil Microscopic observation lymhocyte transformation rate, mtt assay under inverted microscope and detect cell activation propagation
A 492Value.Wherein, living cells ratio 〉=95%, lymhocyte transformation rate 〉=70%,
A 492Value between 0.85 ~ 1.20, cellular immune function is normal.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104173252A (en) * | 2014-07-29 | 2014-12-03 | 蔡贤芬 | Biological beautifying preparation containing autologous stroma cells |
| CN104931681A (en) * | 2015-07-14 | 2015-09-23 | 广州赛莱拉干细胞科技股份有限公司 | Method for detecting serum biochemical indices or sub-health improvement effect of immune cells |
| CN108354946A (en) * | 2017-08-25 | 2018-08-03 | 肖定璋 | A kind of preparation method and application of human immunity composite factor |
| CN113274407A (en) * | 2021-03-30 | 2021-08-20 | 广东莱迪生物医药研究院有限公司 | Anti-aging cell preparation and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978159A (en) * | 2012-11-22 | 2013-03-20 | 深圳市中美康士生物科技有限公司 | Method for enhancing reproductive capacity of CIK (Cytokine Induced Killer) cell and improving tumor cell killing capacity |
| CN103255106A (en) * | 2012-02-16 | 2013-08-21 | 孙勇 | Autologous hematopoietic stem cell technology used for sub-health treatment |
-
2013
- 2013-07-17 CN CN201310298878.3A patent/CN103385890B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103255106A (en) * | 2012-02-16 | 2013-08-21 | 孙勇 | Autologous hematopoietic stem cell technology used for sub-health treatment |
| CN102978159A (en) * | 2012-11-22 | 2013-03-20 | 深圳市中美康士生物科技有限公司 | Method for enhancing reproductive capacity of CIK (Cytokine Induced Killer) cell and improving tumor cell killing capacity |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104173252A (en) * | 2014-07-29 | 2014-12-03 | 蔡贤芬 | Biological beautifying preparation containing autologous stroma cells |
| CN104931681A (en) * | 2015-07-14 | 2015-09-23 | 广州赛莱拉干细胞科技股份有限公司 | Method for detecting serum biochemical indices or sub-health improvement effect of immune cells |
| CN108354946A (en) * | 2017-08-25 | 2018-08-03 | 肖定璋 | A kind of preparation method and application of human immunity composite factor |
| CN113274407A (en) * | 2021-03-30 | 2021-08-20 | 广东莱迪生物医药研究院有限公司 | Anti-aging cell preparation and preparation method thereof |
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| CN103385890B (en) | 2015-09-23 |
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Effective date of registration: 20190531 Address after: 050081 No. 450 Zhongshan West Road, Shijiazhuang City, Hebei Province Co-patentee after: Cui Cheng Patentee after: Chinese people's Liberation Army Army Military Medical University Sergeant school Address before: 050081 No. 450 Zhongshan West Road, Shijiazhuang City, Hebei Province Patentee before: Cui Cheng |
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Effective date of registration: 20200703 Address after: 050081 No. 450 West Zhongshan Road, Hebei, Shijiazhuang Patentee after: NONCOMMISSIONED OFFICER ACADEMY OF ARMY MEDICAL University Address before: 050081 No. 450 West Zhongshan Road, Hebei, Shijiazhuang Co-patentee before: Cui Cheng Patentee before: NONCOMMISSIONED OFFICER ACADEMY OF ARMY MEDICAL University |
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