Multi-cell extract, preparation method and application of multi-cell extract in anti-aging skin care products

Abstract

The invention relates to the technical field of skin care products, in particular to a multi-cell extract, a preparation method and application of the multi-cell extract in anti-aging skin care products. The multi-cell extract comprises a leukocyte extract and at least one plant cell extract, the plant cell extract is derived from any one or a mixture of more of MALUS DOMESTICA fruit cell culture extract and ADENIUM OBESUM leaf cell extract, each component of the multi-cell extract is derived from natural plants and leukocytes, and each component of the multi-cell extract is derived from natural plants and leukocytes. The traditional Chinese medicine composition is safe and reliable, small in irritation to skin, simple in component composition, lower in cost and low in sensitization, and has a good market prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of skin care products, and in particular to a multiple cell extract, a preparation method and application thereof in anti-aging skin care products. Background Art

[0002] Leukocytes are a type of colorless nucleated blood cells, which can be divided into granulocytes, monocytes and lymphocytes according to their morphology, function and source. Leukocyte extract (LEUKOCYTE EXTRACT) is a cell secreted complex extracted from leukocytes. The existing method of extracting leukocytes as raw materials mainly includes lysing leukocytes to obtain leukocyte extracts. The main components of the product obtained by this method are nucleotides and contain extremely small amounts of cytokines or small molecule polypeptides. Its main uses include anti-inflammatory (Wang Bin, Ge Zhidong, Chen Minzhu, et al. Anti-inflammatory effect of leukocyte extract [J]. Journal of Anhui Medical University, 1998, (05): 19-20. DOI:10.19405 / j.cnki.issn1000-1492.1998.05.007.), anti-pulmonary fibrosis (Wang Chunlin, Fan Qixiu, Liu Jiewen, et al. Effect of leukocyte extract on bleomycin-induced experimental pneumonia and pulmonary fibrosis [J]. Chinese Journal of Labor Hygiene and Occupational Diseases, 2001, (02): 48-49), antioxidant and anti-aging effects.

[0003] Leukocyte extract (LEUKOCYTE EXTRACT) is widely used in the field of skin care. This raw material is included in the "Catalogue of Used Cosmetic Raw Materials (2021 Edition)" issued by the State Food and Drug Administration, and its safety is relatively good. At the same time, some patent documents also disclose the use of leukocyte extract as one of the raw materials in skin care products. For example, the Chinese invention patent with document number CN116948961B discloses a leukocyte extract, an anti-wrinkle and firming cream and a preparation method thereof. This invention patent uses animal umbilical cord blood as a raw material, separates and produces leukocyte extract, and then adds it to the cream base liquid containing transdermal ingredients to prepare a leukocyte extract anti-wrinkle and firming cream rich in multiple growth and cytokines. The Chinese invention patent with the document number CN113750031B discloses an antioxidant composition and its preparation method, wherein the antioxidant composition includes leukocyte extract and platelet-rich plasma, wherein the leukocyte extract is rich in nucleotides, and the platelet-rich plasma is rich in various growth factors and has the function of inhibiting oxidative stress. By mixing the two to form a composition, the combined advantages of the two are fully utilized to significantly improve the antioxidant capacity, enhance the antioxidant capacity of fibroblasts, and effectively slow down skin aging. Such technologies have shown good application prospects of leukocyte extracts in the field of skin care.

[0004] At the same time, there are many skin care products containing leukocyte extracts on the market. For example, the ingredients of Hubei G makeup network preparation number 2021000270 include: leukocyte extract, SCUTELLARIA BAICALENSIS root extract, Centella asiatica extract, Rosemary (ROSMARINUS OFFICINALIS) leaf extract, Polygonum cuspidatum root extract, Matricaria (CHAMOMILLA RECUTITA) flower extract, etc.; the main ingredients of the skin care products of Shanghai G makeup network preparation number 2024002004 are leukocyte extract, Dendrobium officinale stem extract, orchid extract, CYCNOCHES COOPERI flower / leaf extract, Helianthus annuus seed oil, Vanilla flat fruit orchid (Vanilla recutita) flower extract, etc. Planifolia fruit extract, Lyciumchinese extract, Gentiana scabra root extract, Zingiber officinale root extract, etc. This type of skin care products uses white blood cell extract as one of the active ingredients to achieve antioxidant and anti-aging effects, but its composition is complex. On the one hand, it makes the raw material cost high, and on the other hand, the complex ingredient system also means that more different chemical ingredients are introduced into skin care products. Each plant extract may contain a variety of active ingredients, such as flavonoids, phenols, terpenes, etc. These ingredients have different chemical structures and different mechanisms of action. When a large number of different chemical ingredients exist in skin care products at the same time, they may interact with each other to produce new compounds or change the properties of the original ingredients. This complex ingredient system increases the chance of skin contact with potential allergens, making it more likely to cause skin allergies in consumers. At the same time, an overly complex ingredient system may also reduce the stability of the product and even lead to antagonism of efficacy. Therefore, how to improve the antioxidant and anti-aging effects of leukocyte extracts and make the skin care product ingredient system as simple as possible has become one of the urgent issues to be solved in order to improve the application of leukocyte extracts in the field of skin care products. Summary of the invention

[0005] In order to solve the above technical problems, the purpose of the present invention is to provide a multiple cell extract, a preparation method and its application in anti-aging skin care products. The multiple cell extract combines leukocyte extract with at least one plant cell to make the multiple cell extract components simple and have excellent anti-aging effects, so that the leukocyte extract has greater application prospects in the field of anti-aging skin care products.

[0006] In order to achieve the above technical effects, the present invention adopts the following technical solutions: First, the present invention provides a multiple cell extract, including a leukocyte extract and at least one plant cell extract.

[0007] The plant cell extract is a mixture of any one or more of an apple (MALUS DOMESTICA) fruit cell culture extract and a desert rose (ADENIUM OBESUM) leaf cell extract.

[0008] The multiple cell extracts include the following cell extracts in parts by weight: 0.5-1.8 parts of leukocyte extract, 20-30 parts of apple (MALUS DOMESTICA) fruit cell culture extract and 3-6 parts of desert rose (ADENIUMOBESUM) leaf cell extract.

[0009] Secondly, the present invention provides a method for preparing a multiple cell extract, wherein the multiple cell extract comprises at least a leukocyte extract, and the method for preparing the multiple cell extract comprises at least the following steps: A1: Obtain healthy umbilical cord blood, which is derived from any one of Siamese crocodile, cow, horse or donkey; A2: The obtained umbilical cord blood is centrifuged to obtain an intermediate layer containing white blood cells through the first centrifugation, and the intermediate layer is aspirated and subjected to secondary centrifugation 2 to 3 times to obtain white blood cells; A3: The leukocytes are fully lysed using a lysis solution to obtain a leukocyte extract.

[0010] Preferably, the centrifugation parameters in A2 are as follows: the first centrifugation condition is 1500-2000 rpm for 10-15 min, and the temperature is controlled at 0-4°C; after the first centrifugation, the middle layer containing white blood cells is taken and PBS buffer is added to suspend it, and the second centrifugation condition is controlled at 2000-2500 rpm, the centrifugation time is 5 min, and the temperature is controlled at 0-4°C. After centrifugation, the supernatant is discarded; after performing the second centrifugation once, the second centrifugation is repeated at least 1-2 times to finally obtain a precipitate, i.e., the washed white blood cells.

[0011] Preferably, the lysate comprises the following components: EDTA, ε-aminocaproic acid and SDS; More preferably, the lysate further comprises any one of dithiothreitol, β-mercaptoethanol, glycerol, Tris and electrolytes.

[0012] Preferably, the lysis solution is composed of: 1-5 mmol / L EDTA, 12-15 mmol / L ε-aminocaproic acid, 1-5 mmol / L dithiothreitol, 10-100 mmol / L β-mercaptoethanol, 4%-6% glycerol, 17-22 mmol / L Tris, 100-120 mmol / L NaCl, 4-6 mmol / L KCl, 0.8%-1.2% SDS (w / v) and proteinase K 0.5 g / L.

[0013] Most preferably, the lysis solution is composed of: 5 mmol / L EDTA, 13 mmol / L ε-aminocaproic acid, 2 mmol / L dithiothreitol, 60 mmol / L β-mercaptoethanol, 5% glycerol, 20 mmol / L Tris, 110 mmol / L NaCl, 4 mmol / L KCl, 1% SDS (w / v) and proteinase K 0.5 g / L.

[0014] In a third aspect, the present invention also provides a use of the aforementioned multiple cell extracts in anti-aging skin care products.

[0015] Preferably, the skin care product is a leave-on skin care product, and the amount of the multiple cell extract added to the leave-on skin care product is 3 to 8 wt %.

[0016] Furthermore, the preparation method of the skin care product is as follows: S1: providing a liquid matrix, wherein the liquid matrix is ​​water or an aqueous solution; preferably, the aqueous solution is ROSA DAMASCENA floral water; S2: While stirring, slowly add the apple (MALUS DOMESTICA) fruit cell culture extract, the leukocyte extract and the desert rose (ADENIUM OBESUM) leaf cell extract, and continue stirring to ensure that each extract is fully dispersed in the liquid matrix to obtain a mixed system containing multiple cell extracts; S3: Adding other formula auxiliary materials to the mixed system, wherein the formula auxiliary materials are any one or more of flavors and fragrances, colorants, pH adjusters, preservatives, penetration enhancers, emulsifiers, thickeners or other functional ingredients to prepare the skin care product.

[0017] It should be noted that the flavors and fragrances, colorants, pH adjusters, preservatives, penetration enhancers, emulsifiers, and thickeners can be arbitrarily selected from the prior art, and the present invention does not impose any limitation thereto.

[0018] Compared with the prior art, the present invention has the following beneficial effects: First, the present invention provides a multi-cell extract that combines a leukocyte extract with at least one plant cell extract and applies it to the field of anti-aging skin care products. In the multi-cell extract, its components are derived from natural plants and leukocytes, which are relatively safe and reliable, less irritating to the skin, and the component categories are simple, the cost is lower, and the sensitization is also low. In the multi-cell extract, the leukocyte extract may contain some growth factors and immunomodulatory factors, which can stimulate the regeneration and repair of skin cells, which helps to replace damaged skin cells and make the skin healthier and more vibrant; and the apple (MALUS DOMESTICA) fruit cell culture extract is rich in a variety of nutrients, such as vitamins, minerals and antioxidants, which can provide nutrients for skin cells, promote cell metabolism and regeneration, thereby reducing the appearance of fine lines and wrinkles; Desert Rose (ADENIUM OBESUM) leaf cell extract may also have the effect of promoting cell regeneration. It may contain some special bioactive ingredients that can stimulate the proliferation and differentiation of skin cells and accelerate the skin repair process. The aforementioned active ingredients all have certain anti-aging effects when used alone, and in a new multiple cell extract provided by the present invention, the combination of the leukocyte extract and the plant cell extract further produces a synergistic effect, thereby enhancing the anti-aging effect of the composition.

[0019] In addition, the inventors of the present invention have further improved the preparation method of leukocyte extracts. The improved extraction method showed better efficacy test results in multiple cell extracts. The inventors speculate that the possible principles are as follows: on the one hand, the lysis method can more effectively release the active ingredients in leukocytes, such as growth factors, cytokines and immunomodulatory factors; on the other hand, by adding protective agents, the damage to the active ingredients in leukocytes during the lysis process is reduced, so that they maintain higher biological activity in skin care products. Taking these two factors into consideration, the appropriate lysis solution composition and ratio have a greater impact on the lysis effect. DETAILED DESCRIPTION

[0020] The following examples are only used to more clearly illustrate the technical scheme of the present invention, and are therefore only used as examples, and cannot be used to limit the scope of the present invention. For those skilled in the art, any equivalent modifications and substitutions to the embodiments described below are also within the scope of the present invention. Therefore, the equalization transformations and modifications made without departing from the spirit and scope of the present invention should all be included in the scope of the present invention. Those who do not specify specific conditions in the embodiments are carried out according to normal conditions or the conditions recommended by the manufacturer. All reagents or instruments that do not specify manufacturers are conventional products that can be purchased commercially.

[0021] In order to better illustrate the present invention, numerous specific details are provided in the following specific embodiments. It should be understood by those skilled in the art that the present invention can be implemented without certain specific details. In other embodiments, methods, means, equipment and steps well known to those skilled in the art are not described in detail in order to highlight the subject matter of the present invention.

[0022] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art. Unless otherwise specified, the units used in this specification are all international standard units, and the numerical values ​​and numerical ranges appearing in the present invention should be understood to include the inevitable errors in industrial production.

[0023] It should be noted that: generally, the nomenclature and its techniques used in conjunction with cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization described herein are those well-known and commonly used in the art. Unless otherwise indicated, the methods and techniques of the present invention are generally carried out according to conventional methods as are well known in the art and as described in various general and more specific references, which are cited and discussed throughout this specification. Enzymatic reactions and purification techniques are carried out according to the manufacturer's instructions, as are commonly implemented in the art, or as described herein. The nomenclature and its laboratory procedures and techniques used in conjunction with analytical chemistry, synthetic organic chemistry, and medical and pharmaceutical chemistry described herein are those well known and commonly used in the art. Example 1

[0024] This embodiment provides a method for preparing a leukocyte extract and a leukocyte extract prepared by the method, specifically: The preparation method of the leukocyte extract is as follows: A1: Obtain healthy umbilical cord blood, which comes from healthy cows that have been tested for viruses (this sample is used in all the following examples); A2: The umbilical cord blood was centrifuged to obtain the intermediate layer containing white blood cells after the first centrifugation. The parameters of the first centrifugation were: speed 2000 rpm, centrifugation 10 min, temperature control at 4°C; After the first centrifugation, the middle layer containing leukocytes was aspirated and PBS buffer was added to suspend the leukocytes, and then a second centrifugation was performed. The conditions of the second centrifugation were controlled at 2500 rpm, the centrifugation time was 5 min, and the temperature was controlled at 4°C. After centrifugation, the supernatant was discarded to obtain the precipitate, and PBS buffer was added to the precipitate and the second centrifugation was repeated twice under the above conditions. The final precipitate obtained was the washed leukocytes.

[0025] A3: Using a lysis solution to fully lyse the leukocytes to obtain a leukocyte extract. Specifically, the operation method is as follows: First, the lysis solution was prepared according to the following composition: 5 mmol / L EDTA, 13 mmol / L ε-aminocaproic acid, 2 mmol / L dithiothreitol, 60 mmol / L β-mercaptoethanol, 5% glycerol, 20 mmol / L Tris, 110 mmol / L NaCl, 4mmol / L KCl, 1% SDS (w / v) and proteinase K 0.5 g / L. The specific operation was carried out in a fume hood, and proteinase K was added after the other components during the preparation process. The finished lysis solution was recorded as lysis solution I.

[0026] Add 500 uL of the prepared lysis solution I to the washed leukocytes (precipitate in step A2), gently invert the centrifuge tube to fully mix the leukocytes and lysis solution, and place in a 37°C water bath for 15 min. During the lysis process, invert the centrifuge tube every 5 min to ensure sufficient lysis. After lysis, obtain a leukocyte mixture.

[0027] The lysed leukocyte mixture was centrifuged in a low-temperature centrifuge at 12,000 rpm, 6 min, and 4°C. The supernatant was taken out after centrifugation to obtain the leukocyte extract. The leukocyte extract obtained by this experimental operation was recorded as leukocyte extract I. Test Example 1

[0028] This embodiment provides a composition comprising multiple cell extracts, in which the multiple cell extracts include leukocyte extracts and at least one plant cell extract, and the composition of the multiple cell extracts in each composition is different. This test example provides the following multiple compositions, and the composition of each composition is shown in Table 1.

[0029] Table 1 Compositions of Composition A to Composition I (parts by mass)

[0030] It should be noted that the sources of the raw materials in the above Table 1 are: the leukocyte extract is the "leukocyte extract I" prepared according to the method of Example 1, and the remaining raw materials, apple (MALUS DOMESTICA) fruit cell culture extract, desert rose (ADENIUM OBESUM) leaf cell extract, alpine edelweiss (LEONTOPODIUM ALPINUM) callus extract, sea fennel (CRITHMUM MARITIMUM) callus culture, myrrh (COMMIPHORA MYRRHA) leaf cell extract, grape (VITIS VINIFERA) flower cell extract, and Rosa DAMASCENA flower water are all commercially available products.

[0031] This test example also includes performing efficacy tests on the compositions A to I prepared above, specifically including: DPPH clearance test DPPH is a stable free radical. When antioxidants are present, they can react with DPPH, changing the color of DPPH from dark purple to light yellow. The scavenging rate of DPPH free radicals can be calculated by measuring the change in absorbance. The higher the DPPH scavenging rate, the stronger the ability of the substance to scavenge DPPH free radicals, and the better the antioxidant effect. DPPH scavenging rate is one of the commonly used methods for evaluating antioxidants in this field.

[0032] The specific experimental method is as follows: prepare a 0.1 mM DPPH working solution and determine its blank absorbance value; then take 100 uL of each of the prepared composition A to composition I solutions and add them to the prepared 2 mL DPPH working solution, react in the dark for 30 minutes, and then determine the absorbance value of each sample after the reaction. The free radical scavenging rate is calculated based on the above two absorbances. Each group of experiments is repeated three times. The experimental results are shown in Table 2. Table 2 DPPH clearance test results of composition A to composition I (n=3, x±s, %) DPPH free radical scavenging rate Composition A 32.51±3.25 Composition B 76.24±4.69 Composition C 41.36±2.34 Composition D 52.15±2.64 Composition E 68.91±3.27 Composition F 43.34±2.94 Composition G 54.35±3.71 Composition H 46.25±3.94 Composition I 56.45±3.31 The above experimental results show that the best combination of antioxidant effects among the compositions provided by the present invention (compositions A to I) is composition B, whose corresponding combination is a combination of leukocyte extract, apple (MALUS DOMESTICA) fruit cell culture extract and desert rose (ADENIUM OBESUM) leaf cell extract; the second is composition E, whose corresponding combination of multiple cell extracts is a combination of leukocyte extract, apple (MALUS DOMESTICA) fruit cell culture extract, desert rose (ADENIUM OBESUM) leaf cell extract and alpine edelweiss (LEONTOPODIUM ALPINUM) callus extract. The experimental results show that the components of the multiple cell extracts in composition B have a good synergistic effect.

[0033] 1.2 Effects of each composition on the proliferation rate of human fibroblasts and the synthesis rate of type I collagen The experimental method is as follows: after the purchased fibroblasts are revived, they are incubated in a cell culture incubator (37°C, 5% CO2) using DMEM medium containing 10% fetal bovine serum (FBS). When the cells grow to a confluence of more than 80%, they are digested with trypsin and passaged. After the fibroblasts in the logarithmic growth phase are digested, they are inoculated in a 96-well culture plate, and 100 uL of cell suspension is added to each well (cell suspension: 1*10 5 cells), so that the cells are evenly distributed, and 10uL of composition A to composition I solution is added to each well; After incubation for 24 hours, the cell proliferation rate was determined by the WST-8 method. At the same time, the type I collagen content in each well was detected using a type I collagen detection kit. The blank group (without any antioxidant active ingredients) was used as a control. The proliferation rate and type I collagen synthesis rate of each group relative to the blank group were calculated. The experimental results are shown in Table 3: Table 3 Effects of composition A to composition I on fibroblast proliferation rate and type I collagen synthesis rate (n=3, x±s, %) Cell proliferation rate (%) Type I collagen synthesis rate (%) Composition A 108.31±3.34 102.26±4.19 Composition B 138.34±5.11 136.31±6.35 Composition C 113.67±3.45 110.02±4.35 Composition D 118.64±2.92 112.35±3.16 Composition E 126.35±4.02 121.35±4.02 Composition F 116.21±2.37 114.34±5.31 Composition G 118.43±3.02 113.39±3.64 Composition H 115.82±4.10 106.35±2.01 Composition I 119.19±3.79 105.66±4.25 The above experimental results show that composition B has the best effect on the proliferation rate of fibroblasts and the synthesis rate of type I collagen, showing a good synergistic effect among the components. Example 2

[0034] This embodiment provides a plurality of methods for preparing leukocyte extracts and leukocyte extracts prepared by the methods, specifically: This example provides 4 control groups, namely control group I to control group IV. The preparation method of the leukocyte extract prepared in each control group refers to Example 1, and only the composition of the lysate is different from the lysate I used in the preparation process of experimental group I. The composition of the lysate used in each control group is shown in Table 4: Table 4 Composition of lysate used in control group I to control group IV Lysis solutions were prepared according to the compositions in the above table, and leukocytes were lysed using the self-prepared lysis solutions (the rest of the steps were referred to Example 1). The leukocyte extracts finally obtained from control group I to control group IV were recorded as control (EBC) I to control (EBC) IV, respectively. Test Example 2

[0035] On the basis of Example 2, the present invention tests the effects of the leukocyte extract prepared in Example 1 and the leukocyte extracts prepared in control groups I to IV to observe the effects of different leukocyte lysates on the efficacy of the leukocyte extracts. The experimental results are expressed as the test results of the DPPH clearance rate of the composition (1 part by weight of leukocyte extract + 999 parts by weight of ROSA DAMASCENA floral water). The experimental method refers to Experimental Example 1. The results of this experimental example are shown in Table 5: Table 5 Test results of DPPH removal rate of each composition (n=3, x±s, %) Composition composition (mass ratio 1:999) DPPH free radical scavenging rate (%) Composition B Leukocyte extract I + ROSA DAMASCENA flower water 77.19±3.36 Composition 1 Control (EBC) I + Rosa Damascena floral water 69.35±2.69 Composition 2 Control (EBC) II + Rosa Damascena floral water 71.33±3.06 Composition 3 Control (EBC) III + Rosa Damascena floral water 59.12±2.91 Composition 4 Control (EBC) IV + Rosa Damascena floral water 68.24±2.77 The above experimental results show that different leukocyte extract preparation methods (mainly the composition of leukocyte lysate) have a significant impact on the efficacy of leukocyte extract. By adding reducing agents, different compositions of reducing agents, or protective agents to the lysate system, the leukocytes can be fully lysed while retaining the effects of their effective components to a greater extent, thereby improving the anti-aging and antioxidant effects of leukocyte extracts in actual applications.

[0036] The above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, which should be included in the scope of the claims of the present invention.

Claims

1. A multiple cell extract, characterized in that The multiple cell extracts include the following cell extracts in parts by weight: 0.5-1.8 parts of leukocyte extract, 20-30 parts of apple (MALUS DOMESTICA) fruit cell culture extract and 3-6 parts of desert rose (ADENIUM OBESUM) leaf cell extract.

2. The method for preparing a multiple cell extract according to claim 1, characterized in that: The multiple cell extracts at least include a leukocyte extract, and the preparation method of the multiple cell extracts at least includes the following steps: A1: Obtain healthy umbilical cord blood, which is derived from any one of Siamese crocodile, cow, horse or donkey; A2: The obtained umbilical cord blood is centrifuged to obtain an intermediate layer containing white blood cells through the first centrifugation, and the intermediate layer is aspirated and subjected to secondary centrifugation 2 to 3 times to obtain white blood cells; A3: The leukocytes are fully lysed using a lysis solution to obtain a leukocyte extract.

3. The method for preparing a multiple cell extract according to claim 2, characterized in that: The centrifugation parameters in A2 are as follows: the first centrifugation condition is 1500-2000 rpm for 10-15 min, and the temperature is controlled at 0-4°C; after the first centrifugation, the middle layer containing white blood cells is taken and PBS buffer is added to suspend it, and the second centrifugation condition is controlled at 2000-2500 rpm, the centrifugation time is 5 min, and the temperature is controlled at 0-4°C. After centrifugation, the supernatant is discarded; after performing the second centrifugation once, the second centrifugation is repeated at least 1-2 times to finally obtain a precipitate, i.e., the washed white blood cells.

4. The method for preparing a multiple cell extract according to claim 2, characterized in that: The lysing solution comprises the following components: EDTA, ε-aminocaproic acid and any one of SDS, dithiothreitol, β-mercaptoethanol, glycerol, Tris and electrolytes.

5. The method for preparing a multiple cell extract according to claim 4, characterized in that: The lysis solution is composed of: 1-5 mmol / L EDTA, 12-15 mmol / L ε-aminocaproic acid, 1-5 mmol / L dithiothreitol, 10-100 mmol / L β-mercaptoethanol, 4%-6% glycerol, 17-22 mmol / L Tris, 100-120 mmol / L NaCl, 4-6 mmol / L KCl, 0.8%-1.2% SDS (w / v) and proteinase K 0.5 g / L.

6. Use of the multiple cell extract as claimed in claim 1 in anti-aging skin care products.

7. The use according to claim 6, characterized in that The skin care product is a leave-on skin care product, and the added amount of the multiple cell extract in the leave-on skin care product is 3 to 8 wt %.

8. The use according to claim 7, characterized in that The preparation method of the skin care product is as follows: S1: providing a liquid matrix, wherein the liquid matrix is ​​water or an aqueous solution; S2: While stirring, slowly add the apple (MALUS DOMESTICA) fruit cell culture extract, the leukocyte extract and the desert rose (ADENIUM OBESUM) leaf cell extract, and continue stirring to ensure that each extract is fully dispersed in the liquid matrix to obtain a mixed system containing multiple cell extracts; S3: Adding other formula auxiliary materials to the mixed system, wherein the formula auxiliary materials are any one or more of flavors and fragrances, colorants, pH adjusters, preservatives, penetration enhancers, emulsifiers, thickeners or other functional ingredients to prepare the skin care product.

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