CN106074356A - A kind of preparation method and applications of living cells essence - Google Patents
A kind of preparation method and applications of living cells essence Download PDFInfo
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Abstract
The invention discloses the preparation method and applications of a kind of living cells essence, the preparation method of described living cells essence, including step: A) use the mescenchymal stem cell in mescenchymal stem cell culture medium mescenchymal stem cell that umbilical cord tissue originate or placenta tissue source to cultivate, collection cell culture fluid；B) to step A) gained cell culture fluid adds pure rose flower water, obtain living cells mixed liquor.The preparation method of living cells essence of the present invention has broken away from traditional plant component extraction and fine chemistry industry production technology, use the method for cell engineering, obtain the Mesenchymal stem cell nutrient solution of the beneficial activity protein molecular containing parent's human body skin by cultivating the mescenchymal stem cell in human umbilical tissue source, using gained cell culture fluid as the basic components of whole product mesenchymal stem cells mixed liquor, and without chemical industry additive in mescenchymal stem cell mixed liquor, at utmost ensure that the safety that product persistently uses.
Description
Technical field
The invention belongs to biological technical field, relate to the preparation method and applications of a kind of living cells essence.
Background technology
The main cause of skin aging is skin dehydration and sebum peroxidating.Along with the increase at age, smegma by
The fewest, add the ill effect of Exposure to Sunlight and free radical, epidermis is the most thinning, and epidermis swells, and mammillary body is flat, and protein generates
Reducing, subcutaneous fat reduces, and easily causes sagging or wrinkle, and the colour of skin is dark and gloomy；On the other hand, the blood flow circulation decline of skin, blood
Pipe resistance dies down, and increasingly generates speckle, causes the aging of skin.
Along with growth in the living standard, the people are to beautiful pursuit, and daily life skin protection is paid attention to by especially women further.From
Initially cleaning and effect of protection skin, till now with the cosmetics that physiological function is the most particularly customized, high security and more
Science, will be following development trend.
Summary of the invention
In consideration of it, the technical problem to be solved in the present invention is to provide a kind of living cells mixed liquor and preparation method thereof, U.S.
Holding preparation, this cosmetic formulation effect is preferable.
The invention provides the living cells essence of a kind of anti-ageing whitening, acceptable auxiliary by effective ingredient and beauty treatment fields
Material composition, described effective ingredient includes that human mesenchymal stem cell cytokine, described adjuvant include that arbutin, trehalose, dimension are raw
Element E and glycerol, step is as follows:
(1) mescenchymal stem cell using mescenchymal stem cell culture medium to originate umbilical cord tissue is cultivated, and collects cell training
Nutrient solution；
(2) (pure rose flower water comes to add the pure rose flower water that volume ratio is 20%--40% in step (1) gained cell culture fluid
From Lv Xiang island, Guangzhou article of everyday use company limited, lot number N020160418), obtain living cells mixed liquor.
Described human stem cell growth is secreting type human stem cell growth, including basic fibroblast growth because of
Son (bFGF), keratinocyte growth factor (KGF), transforming growth factor-beta 1 (TGF-β 1), VEGF (VEGF)
With at least two in insulin like growth factor (IGF).
Human stem cell growth concentration is 1～40mg/ml.
Wherein preferred version is: the concentration of keratinocyte growth factor is 10mg/ml, basic fibroblast growth factor
Concentration be 8mg/ml, the concentration of transforming growth factor-beta 1 is 7mg/ml, and the concentration of VEGF is 10mg/ml,
The concentration of insulin like growth factor is 5mg/ml；
Further: in step (2) gained cell culture fluid, also to add arbutin, trehalose, vitamin E and glycerol.
In described mescenchymal stem cell mixed liquor: the concentration of arbutin is 1-5mg/ml；The concentration of trehalose is 1-
5mg/ml；The concentration of vitamin E is 1-5mg/ml, and the concentration of glycerol is 5-10mg/ml.
The wherein preparation of living cells mixed liquor, more specifically step is:
(1) gather and obtain neonatal umbilical cord mescenchymal stem cell: with the normal saline flushing umbilical cord containing mycillin
Band tissue, is layered in Tissue Culture Dish cultivation after being cut into small pieces；
(2) Tissue Culture Dish poly-D-lysine is coated, and uses after naturally drying；
(3) with primary culture medium to adherent stem cell being carried out original cuiture, a cell culture fluid, digestion within every three days, are changed
Cell, digestion terminates rear Secondary Culture base and carries out terminating digestion, passes in adherent area 1:3 ratio；
(4) collect cell culture fluid, collect the cell culture supernatant before passing on, obtain cell culture fluid；
(5) cell culture fluid collected, through filtration system, filters off cell debris and microorganism；
(6) gained culture fluid mixes with MEIGUI CHUNLU, i.e. obtains described living cells mixed liquor；It is placed in 0-8 DEG C of storage.
The mescenchymal stem cell mixed liquor prepared by said method, can be widely applied in cosmetics.
Compared with prior art, there is advantages that
Cell growth factor plays physiological regulatory action to various kinds of cell physiological function and metabolic activity, they content poles in vivo
Micro-, but biological activity is high.In beauty and skin care, plastic surgery, burn ulcer and various dermopathic wound repair and healing
In play an important role.
Rich in basic fibroblast growth factor (bFGF) in cytokine, keratinocyte growth factor (KGF), convert growth
The factor-β 1(TGF-β 1), VEGF (VEGF) and insulin like growth factor (IGF) and other multiple nutrients thing
Matter, effect is:
1, nutrition is provided: containing skin daily need, enough nutritional labeling in elite, supplement the most moist to skin；
2, epidermal growth factor combine with the horn cell factor, insulin like growth factor arbutin promote skin epidermal area and
Horny layer hypertrophy is broken up, and strengthens thickness and the elasticity of skin；
3, VEGF can promote the blood supply of superficial cutaneous capillaries, regulates the poised state on shallow top layer, improves skin
Hydropenia situation, plays the effect of lock water moisturizing；
4, fibroblast growth factor promotes formation and the filling of collagen protein of skin fiber, makes skin more glossy；
5, effectively suppress melanic formation, subtract light cicatrix, delay skin aging.
This product, without essence, because the skin care item of major part factor-containing have the slight smell of blood, uses Flos Rosae Rugosae pure
Dew can hide the smell of blood, also whitening function.
The consumption of each raw material of the present invention and physicochemical property produce coordinative role, have the active effect that repair cell, beautiful
Bai Yangyan.PH value is close with the pH value of human body skin, has no stimulation skin.Substantially feel after using to moisten, soft, oil-free
Soapy feeling.There is obvious whitening to skin moisten, the effect of cosmetology.
Mescenchymal stem cell mixed liquor of the present invention has broken away from traditional plant component extraction and fine chemistry industry production technology, adopts
Mescenchymal stem cell by the method for cell engineering, by cultivation human umbilical tissue source obtains and contains the useful of parent's human body skin
The Mesenchymal stem cell nutrient solution of activated protein molecule, using gained cell culture fluid as whole product mesenchymal stem cells mixed liquor
Basic components, and without chemical industry additive in mescenchymal stem cell mixed liquor, such as: ethanol, lead hydrargyrum, coloring agent, mineral
The sensitization toxin such as oil, at utmost ensure that the safety that product persistently uses.
The present invention adds arbutin and vitamin E in mescenchymal stem cell mixed liquor, and trace arbutin can stimulate cell
The speed of growth speeds, and improves dermal layer of the skin histiocyte and fibroblastic division growth power；Vitamin E can allow arbutin
In essence more stable, the most crease-resistant, aging resistance effect；The present invention contains alkalescence fibroblast in mescenchymal stem cell mixed liquor
Dimension cell growth factor, keratinocyte growth factor, transforming growth factor-beta 1, these several somatomedin have strengthening skin screen
Hinder defensive enginery, to free radical resisting antioxidative effect；Active factors therein includes VEGF, Insulin-Like
Somatomedin, the whitening and moisturizing reparation for skin has more preferable effect.The present invention is also especially added with trehalose, and it can promote
Produce, strengthen the defensive enginery of skin keratin confluent monolayer cells environment to external world in entering skin at collagen and elastic fiber；Between the present invention
Various formula components in mesenchymal stem cells mixed liquor are worked in coordination with mutually, are achieved its optimal skin care effect.
The preparation method of mescenchymal stem cell mixed liquor of the present invention is simple, can carry out large-scale promotion and form industry metaplasia
Produce processing.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described in further detail:
(1) mescenchymal stem cell using mescenchymal stem cell culture medium to originate umbilical cord tissue is cultivated, and collects cell training
Nutrient solution；
(2) (pure rose flower water comes to add the pure rose flower water that volume ratio is 30% in step (1) gained cell culture fluid
From Lv Xiang island, Guangzhou article of everyday use company limited, lot number N020160418), obtain living cells mixed liquor.
Further: in step (2) gained cell culture fluid, also to add arbutin, trehalose, vitamin E and glycerol.
Step one: arbutin, vitamin E are dispersed in glycerol, is then heated to 50 DEG C by this mixture, all
Dissolve；
Step 2: stem cell mixed liquor is heated to 37 DEG C of defrostings；
Step one material is slowly added in the material to step 2, cooling discharging after stirring.
Described living cells essence, raw material in its formula and weight ratio be respectively: human stem cell growth concentration
For 40mg/ml；The concentration of arbutin is 5mg/ml；The concentration of trehalose is 5mg/ml；The concentration of vitamin E is 5mg/ml, sweet
The concentration of oil is 10mg/ml.
Contrast test
Choose 30 25 55 years old healthy, without dermopathic women, be randomly divided into experimental group and matched group；Examination is made before test
Test group and the volunteer of matched group under conditions of room temperature 20-25 DEG C, humidity 40-60%, within 1 hour in advance, enter in test environment
Quiet to be measured, the left and right forearm at test object determines test zone and exposes for 20 minutes in advance, area about 30mm ×
30mm；
Use TMT210Tewameter moisture of skin scatter and disappear tester carry out electric capacity and moisture through skin scatter and disappear rate test, use
SEM575 type skin elasticity detector carries out skin elasticity and compactibility detection, and test result is initial value；
Test group uses living cells essence, and matched group uses 10% sodium hyaluronate solution；Using method is divided into the most each painting
Smear the same area once, use 28 days continuously.After in test, the 28th day same day used test specimen and reference substance for the last time, make
The volunteer of test group and matched group keeps 1 hour in room temperature 20-25 DEG C, the environment of humidity 40-60%, measures first 20 minutes
Exposure is put to the test domain test, and test result is test value；
Matched group test value compares with initial value, and the moisture of skin rate of scattering and disappearing of matched group adds 14.29%；Average capacitance
Add 1.8%；Skin elasticity increases by 6.3%；The average compactibility of skin increases by 0.2%；
Test group test value compares with initial value, and the moisture of skin rate of scattering and disappearing of test group adds 50%；Average capacitance increases
11%；Skin elasticity increases by 0.4%；The average compactibility of skin increases by 4.5%.
Matched group result of the test:
| Project | TEWL value (g/m2h) | Average capacitance (u.S.) | Average elasticity (%) | Average compactibility (%) |
| Initial value | 14 | 223.7 | 74.3 | 69.7 |
| Test value | 12 | 227.9 | 76.5 | 70.5 |
| Increment rate (%) | 14.29 | 1.88 | 2.96 | 1.15 |
Test group result of the test:
| Project | TEWL value (g/m2h) | Average capacitance (u.S.) | Average elasticity (%) | Average compactibility (%) |
| Initial value | 16 | 225.5 | 75.4 | 69.5 |
| Test value | 8 | 250.8 | 80.7 | 74.3 |
| Increment rate | 50 | 11.22 | 7.03 | 6.91 |
The above results shows, living cells essence has the physiologically active being obviously promoted keratinocyte growth and breeding, it is possible to
Repair skin, strengthen compactibility and the elasticity of skin, the effect of opposing skin aging can be produced.Meanwhile, living cells essence
In cytokine contain substantial amounts of polar group, can as Moisture factor, because of and there is the tyrosine stoped in skin
It is converted into melanic effect, makes skin moisture-keeping whitening.
Claims (8)
1. a preparation method for living cells essence, is characterized in that: comprise the following steps:
A) mescenchymal stem cell culture medium mescenchymal stem cell that umbilical cord tissue originate or placenta tissue is used to fill between originating
Matter is cultivated by stem cell, collects cell culture fluid；
B) to step A) gained cell culture fluid adds the pure rose flower water that volume ratio is 20%--40%, obtain living cells
Mixed liquor.
2., required the preparation method of the living cells essence as described in 1, it is characterized in that: the stem cell in described stem cell media
Somatomedin is secreting type stem cell factor, for basic fibroblast growth factor, keratinocyte growth factor, converts
At least two in growth factor-beta 1, VEGF and insulin like growth factor.
3. the preparation method of living cells essence as claimed in claim 2, is characterized in that: stem cell factor concentration is 1
～40mg/ml.
4. the preparation method of living cells essence as claimed in claim 3, is characterized in that: described stem cell factor includes
5 kinds, wherein the concentration of keratinocyte growth factor is 10mg/ml, and the concentration of basic fibroblast growth factor is 8mg/ml,
The concentration of transforming growth factor-beta 1 is 7mg/ml, and the concentration of VEGF is 10mg/ml, insulin-like growth factor
The concentration of son is 5mg/ml.
5. the preparation method of living cells essence as claimed in claim 1, is characterized in that: further comprise the steps of:
C) to step B) gained cell culture fluid adds arbutin, trehalose, vitamin E and glycerol.
6. the preparation method of living cells essence as claimed in claim 5, is characterized in that: the concentration of described arbutin is 1-
5mg/ml, the concentration of trehalose is 1-5mg/ml, and the concentration of vitamin E is 1-5mg/ml, and the concentration of glycerol is 5-10mg/ml.
The preparation method of a kind of living cells mixed liquor the most according to claim 1, it is characterised in that:
Described A) step includes:
A1) gather and obtain neonatal umbilical cord mescenchymal stem cell: with the normal saline flushing umbilical cord containing mycillin
Band tissue, is layered in Tissue Culture Dish cultivation after being cut into small pieces；
A2) Tissue Culture Dish poly-D-lysine is coated, and uses after naturally drying；
A3) with primary culture medium to adherent stem cell being carried out original cuiture, a cell culture fluid, digestion within every three days, are changed
Cell, digestion terminates rear Secondary Culture base and carries out terminating digestion, passes in adherent area 1:3 ratio；
A4) collect cell culture fluid, collect the cell culture supernatant before passing on, obtain cell culture fluid；
A5) cell culture fluid collected, through filtration system, filters off cell debris and microorganism.
8. a cosmetic formulation, is characterized in that, described cosmetic formulation includes prepared by preparation method described in any one of claim 1-7
And the living cells mixed liquor come.
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Cited By (7)
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| CN107260652A (en) * | 2017-05-11 | 2017-10-20 | 珠海伊斯佳科技股份有限公司 | A kind of skin care compositions and methods and its application method |
| CN108158949A (en) * | 2018-03-05 | 2018-06-15 | 贵州泛特尔细胞生物技术有限公司 | A kind of anti-ageing smoothing wrinkle Essence and preparation method thereof |
| CN108354895A (en) * | 2018-04-10 | 2018-08-03 | 北昊干细胞与再生医学研究院有限公司 | Composition, facial mask essence containing stem cell medium and preparation method thereof |
| CN108553405A (en) * | 2018-05-31 | 2018-09-21 | 北昊干细胞与再生医学研究院有限公司 | It releives the composition of maintenance skin, cosmetics and preparation method thereof |
| CN110917111A (en) * | 2019-12-30 | 2020-03-27 | 上海掘谜生物科技有限公司 | Preparation method of mesenchymal stem cell derived liquid and application of mesenchymal stem cell derived liquid in cosmetics |
| CN113559047A (en) * | 2020-04-28 | 2021-10-29 | 上海夏必拓生物科技有限公司 | Preparation method of beauty skin care product and skin care essence containing active factor extract secreted by mesenchymal stem cells |
| CN118319831A (en) * | 2024-06-13 | 2024-07-12 | 广州沙艾生物科技有限公司 | Composition containing mesenchymal stem cell culture and application of composition in preparation of skin care product |
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| CN107260652A (en) * | 2017-05-11 | 2017-10-20 | 珠海伊斯佳科技股份有限公司 | A kind of skin care compositions and methods and its application method |
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| CN108553405A (en) * | 2018-05-31 | 2018-09-21 | 北昊干细胞与再生医学研究院有限公司 | It releives the composition of maintenance skin, cosmetics and preparation method thereof |
| CN110917111A (en) * | 2019-12-30 | 2020-03-27 | 上海掘谜生物科技有限公司 | Preparation method of mesenchymal stem cell derived liquid and application of mesenchymal stem cell derived liquid in cosmetics |
| CN113559047A (en) * | 2020-04-28 | 2021-10-29 | 上海夏必拓生物科技有限公司 | Preparation method of beauty skin care product and skin care essence containing active factor extract secreted by mesenchymal stem cells |
| CN118319831A (en) * | 2024-06-13 | 2024-07-12 | 广州沙艾生物科技有限公司 | Composition containing mesenchymal stem cell culture and application of composition in preparation of skin care product |
| CN118319831B (en) * | 2024-06-13 | 2024-09-20 | 广州沙艾生物科技有限公司 | Composition containing mesenchymal stem cell culture and application of composition in preparation of skin care product |
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