Placenta / umbilical cord mesenchymal stem cell culture composition and application thereof in functional anti-aging

Abstract

The invention discloses a placenta / umbilical cord mesenchymal stem cell culture composition and application thereof in functional anti-aging. The stem cell culture composition is culture supernate obtained after mesenchymal stem cells are subjected to induction culture through a first induction culture medium and a second induction culture medium. The first induction culture medium contains 10 to 30 mg / L of rose willow flavone, 5.0 to 7.5 [mu] g / L of transferrin, 0.4 to 1.2 [mu] g / L of bovine serum albumin and 2.0 to 5.5 [mu] g / L of linoleic acid. The second induction culture medium contains 10 to 30 mg / L of rose willow flavone, 60 to 90 mg / L of tetramethylpyrazine, 5.0 to 7.5 [mu] g / L of transferrin, 0.4 to 1.2 [mu] g / L of bovine serum albumin and 2.0 to 5.5 [mu] g / L of linoleic acid. According to the stem cell culture composition obtained through induction of the first induction culture medium containing the salix ramosissima flavone and the second induction culture medium containing the salix ramosissima flavone and the tetramethylpyrazine, the salix ramosissima flavone and the tetramethylpyrazine in the induction culture mediums have a synergistic effect, and generation of some active factors of the stem cells is promoted; the traditional Chinese medicine composition has remarkable effects of resisting oxidation, inhibiting tyrosine, promoting collagen regeneration and the like, has a very prominent anti-aging skin repairing effect, and can be used for brightening the skin color.

Description

Technical Field

[0001] The present invention relates to the technical field of stem cells, in particular to a placenta / umbilical cord mesenchymal stem cell culture composition and its application in functional anti-aging. Background Art

[0002] Mesenchymal stem cells (MSC) are a type of multipotent stem cells that have all the common characteristics of stem cells, namely, self-renewal and multidirectional differentiation capabilities. Mesenchymal stem cell-derived exosomes (MSC-Exo) are nanovesicles responsible for the paracrine effects of MSCs. They contain a variety of contents, including mRNA, miRNA, cytokines, and proteins. They can penetrate the physiological barriers of body tissues and participate in intercellular material exchange and information communication. They have broad application potential in the treatment of diseases such as anti-skin aging, wound repair, scar repair, and nerve regeneration.

[0003] Recently, the research on mesenchymal stem cell-derived exosomes is very popular. Based on the potential of mesenchymal stem cells, this application has developed a mesenchymal stem cell tissue culture, which can promote stem cells to secrete certain active cytokines through induction culture containing a first induction medium containing flavonoids and a second induction medium containing flavonoids and ligustrazine. The active cytokines have significant effects on whitening, anti-oxidation, and promoting collagen regeneration. Summary of the invention

[0004] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a placenta / umbilical cord mesenchymal stem cell culture composition and its application in functional anti-aging to solve the problems raised in the above technical background.

[0005] To achieve the above object, the present invention is implemented through the following technical solutions:

[0006] The present invention provides a placenta / umbilical cord mesenchymal stem cell culture composition, which is a culture supernatant obtained after mesenchymal stem cells are induced and cultured with a first induction medium and a second induction medium;

[0007] The first induction culture medium comprises red safflower flavonoids, transferrin, bovine serum albumin, linoleic acid, and basal culture medium.

[0008] The second induction culture medium comprises flavonoids of red sedge, ligustrazine, transferrin, bovine serum albumin, linoleic acid and basal culture medium.

[0009] Preferably, the first induction culture medium contains 10-30 mg / L flavonoids, 5.0-7.5 ug / L transferrin, 0.4-1.2 ug / L bovine serum albumin, and 2.0-5.5 ug / L linoleic acid.

[0010] Further preferably, the first induction medium contains 20 mg / L flavonoids, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid.

[0011] Preferably, the second induction medium contains 10-30 mg / L flavonoids, 60-90 mg / L chuanxiongzine, 5.0-7.5 ug / L transferrin, 0.4-1.2 ug / L bovine serum albumin, and 2.0-5.5 ug / L linoleic acid.

[0012] Further preferably, the second induction medium contains 20 mg / L flavonoids, 75 mg / L ligustrazine, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid.

[0013] Preferably, the preparation method of the red willow flavonoids is as follows: take dried red willow and grind it with a grinder to pass it through a 40-mesh sieve to obtain red willow powder; weigh the red willow powder, put it in a 250mL round-bottom flask, add 70% volume fraction of ethanol at a solid-liquid ratio of 1g:20ml, and extract at 80°C for 2h; filter while hot, remove ethanol in the filtrate by rotary evaporation, add redistilled water and an equal amount of petroleum ether, extract with a separatory funnel, and discard the petroleum ether layer; extract the water layer twice with 20mL of ethyl acetate, combine the ethyl acetate layers, and rotary evaporate to dryness to obtain red willow flavonoids.

[0014] Preferably, the mesenchymal stem cells are derived from placenta or umbilical cord.

[0015] Preferably, the basal culture medium is DMEM / F12 or α-MEM.

[0016] Preferably, a placenta / umbilical cord mesenchymal stem cell culture composition is prepared by:

[0017] Mesenchymal stem cells were obtained and cultured in a basal medium containing 10% exosome-free fetal bovine serum;

[0018] The cells were transferred to the first induction medium and cultured for 24 hours. When the cell confluence reached 80%, they were transferred to the second induction medium. After continuing to culture for 36 to 48 hours, the cell culture fluid was collected and centrifuged to obtain a mesenchymal stem cell culture composition, which was stored at 4°C for later use.

[0019] In a second aspect, the present invention provides the use of the placenta / umbilical cord mesenchymal stem cell culture composition described above in the preparation of epidermal whitening, skin repair and anti-aging products.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] 1. In the present application, the stem cell culture composition is obtained by induction with a first induction medium containing flavonoids from red safflower and a second induction medium containing flavonoids from red safflower and ligustrazine. The flavonoids from red safflower and ligustrazine in the induction medium act synergistically to promote the production of certain active factors of stem cells, and have significant effects of anti-oxidation, inhibition of tyrosine and promotion of collagen regeneration.

[0022] 2. The stem cell culture composition of the present application has excellent antioxidant, tyrosine inhibition and collagen regeneration promotion effects in in vitro experiments, has a very outstanding anti-aging skin repair effect, and can enhance skin luster and brighten skin tone, and has a good market application prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 A bar graph showing the effect of the stem cell culture composition of the present invention on the tyrosinase inhibition rate;

[0024] Figure 2 The figure is a bar graph showing the effect of the stem cell culture composition of the present invention on the scavenging rate of DPPH free radicals. DETAILED DESCRIPTION

[0025] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0026] The components and reagents involved in the following examples are all conventional commercially available products, such as DMEM / F12 medium and α-MEM medium, which were purchased from Gibco; red willow, which was collected from Karamay, Xinjiang; and chuanxiongzine, which was purchased from Shaanxi Guben Biotechnology Co., Ltd.

[0027] In the embodiments of the present application, mesenchymal stem cells are derived from placenta and umbilical cord; in some embodiments, mesenchymal stem cells can be selected from mammals such as humans, cows, sheep, pigs, rabbits, etc.

[0028] Example 1

[0029] An umbilical cord mesenchymal stem cell culture composition, which is a culture supernatant obtained after umbilical cord mesenchymal stem cells are induced and cultured with a first induction medium and a second induction medium;

[0030] The first induction medium is a DMEM / F12 medium containing 10 mg / L flavonoids, 5.0 ug / L transferrin, 1.2 ug / L bovine serum albumin, and 5.5 ug / L linoleic acid. The second induction medium is a DMEM / F12 medium containing 30 mg / L flavonoids, 60 mg / L ligustrazine, 5.0 ug / L transferrin, 1.2 ug / L bovine serum albumin, and 5.5 ug / L linoleic acid.

[0031] Example 2

[0032] A placental mesenchymal stem cell culture composition, which is a culture supernatant obtained after placental mesenchymal stem cells are induced and cultured with a first induction medium and a second induction medium;

[0033] The first induction medium is a DMEM / F12 medium containing 20 mg / L flavonoids, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid. The second induction medium is a DMEM / F12 medium containing 20 mg / L flavonoids, 75 mg / L ligustrazine, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid.

[0034] Example 3

[0035] A placental mesenchymal stem cell culture composition, which is a culture supernatant obtained after placental mesenchymal stem cells are induced and cultured with a first induction medium and a second induction medium;

[0036] The first induction medium contained 30mg / L flavonoids, 7.5ug / L transferrin, 0.4ug / L bovine serum albumin,

[0037] The second induction medium is α-MEM containing 10 mg / L flavonoids, 90 mg / L ligustrazine, 7.5 ug / L transferrin, 0.4 ug / L bovine serum albumin, and 2.0 ug / L linoleic acid.

[0038] Comparative Example 1

[0039] This comparative example is similar to Example 2, except that: the first induction medium does not contain red safflower flavonoids; specifically: a placental mesenchymal stem cell culture composition, which is a culture supernatant obtained after placental mesenchymal stem cells are induced and cultured in the first induction medium and the second induction medium;

[0040] The first induction medium is a DMEM / F12 medium containing 6.0ug / L transferrin, 0.8ug / L bovine serum albumin, and 3.5ug / L linoleic acid. The second induction medium is a DMEM / F12 medium containing 20mg / L flavonoids, 75mg / L ligustrazine, 6.0ug / L transferrin, 0.8ug / L bovine serum albumin, and 3.5ug / L linoleic acid.

[0041] Comparative Example 2

[0042] This comparative example is similar to Example 2, except that: the second induction medium does not contain red safflower flavonoids; specifically: a placental mesenchymal stem cell culture composition, which is a culture supernatant obtained after placental mesenchymal stem cells are induced and cultured in the first induction medium and the second induction medium;

[0043] The first induction medium is a DMEM / F12 medium containing 20 mg / L flavonoids, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid. The second induction medium is a DMEM / F12 medium containing 75 mg / L ligustrazine, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid.

[0044] Comparative Example 3

[0045] This comparative example is similar to Example 2, except that: the second induction medium does not contain chuanxiongzine; specifically: a placental mesenchymal stem cell culture composition, which is a culture supernatant obtained after placental mesenchymal stem cells are induced and cultured in the first induction medium and the second induction medium;

[0046] The first induction medium is a DMEM / F12 medium containing 20 mg / L flavonoids, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid. The second induction medium is a DMEM / F12 medium containing 20 mg / L flavonoids, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid.

[0047] Comparative Example 4

[0048] This comparative example is similar to Example 2, except that: the second induction medium does not contain flavonoids and ligustrazine; specifically: a placental mesenchymal stem cell culture composition, which is a culture supernatant obtained after placental mesenchymal stem cells are induced and cultured in the first induction medium and the second induction medium;

[0049] The first induction medium is a DMEM / F12 medium containing 20 mg / L flavonoids, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid. The second induction medium is a DMEM / F12 medium containing 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid.

[0050] Comparative Example 5

[0051] A second induction culture medium is a DMEM / F12 culture medium containing 20 mg / L flavonoids, 75 mg / L ligustrazine, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid.

[0052] In the present invention, the preparation method of the red willow flavonoids is as follows: taking dried red willow and crushing it with a grinder to pass through a 40-mesh sieve to obtain red willow powder; weighing the red willow powder, placing it in a 250mL round-bottom flask, adding 70% volume fraction of ethanol at a solid-liquid ratio of 1g:20ml, and extracting it at 80°C for 2h; filtering while hot, removing ethanol in the filtrate by rotary evaporation, adding redistilled water and an equal amount of petroleum ether, extracting with a separatory funnel, and discarding the petroleum ether layer; extracting the water layer twice with 20mL of ethyl acetate, combining the ethyl acetate layers, and rotary evaporating to dryness to obtain red willow flavonoids.

[0053] In the present invention, a mesenchymal stem cell culture composition is prepared according to the formula of Examples 1-3 and Comparative Examples 1-4, wherein the preparation method of the mesenchymal stem cell culture composition is:

[0054] Placenta or umbilical cord mesenchymal stem cells were obtained and cultured in a basal medium containing 10% exosome-free fetal bovine serum, and the cells were collected and cultured in a cell culture incubator at 37°C and 5% CO2 for 48 hours; the basal medium was DMEM / F12 or α-MEM;

[0055] The above cells were divided into 1×10 6 The cells were transferred to the first induction medium at a density of 10 cells / mL for 24 hours. When the cell confluence reached 80%, the cells were transferred to the second induction medium. After 48 hours of continuous culture, the cell culture fluid was collected and centrifuged at 4000 rpm for 20 minutes. The culture fluid was filtered with a 0.22 μm filter to obtain a mesenchymal stem cell culture composition, which was stored at 4°C for later use. Among them, the placental mesenchymal stem cells were rabbit placental mesenchymal stem cells (Cat. No.: CP-Rb303, Specification: 5×10 5 Cells / T25 culture flasks) were purchased from Wuhan Punosai Life Science Co., Ltd.; umbilical cord mesenchymal stem cells were rabbit umbilical cord mesenchymal stem cells (Cat. No.: CP-Rb041, specification: 5×10 5Cells / T25 culture flask) were purchased from Wuhan Pronocell Life Science Technology Co., Ltd.

[0056] To further illustrate the efficacy of the mesenchymal stem cell culture composition provided in the present application example, the present application example also conducted the following experiments:

[0057] 1. Tyrosinase inhibition test

[0058] Tyrosinase is the most critical enzyme in the formation of melanin in the skin, and inhibiting tyrosinase activity can achieve a whitening effect. Therefore, a tyrosinase catalytic reaction system was constructed to evaluate the inhibitory effect of the stem cell culture composition of the present application example on tyrosinase.

[0059] Samples to be tested and grouping: 8 groups are set, which are the stem cell culture compositions or induction culture media provided by Examples 1 to 3 and Comparative Examples 1 to 5 respectively;

[0060] Test method: The inhibition test of each sample to be tested on tyrosinase is carried out according to the tyrosinase catalytic reaction system (Table 1), and each sample is repeated 3 times; the tyrosinase catalytic reaction system is divided into four reaction tubes A1, A2, A3, and A4 for sample addition. After the sample addition, the reaction is carried out at a constant temperature of 37°C in a water bath for 10 minutes, and the absorbance values ​​of A1, A2, A3, and A4 are measured at 475nm. Tyrosinase inhibition rate (%) = [1-(A3-A4) / (A1-A2)] × 100%, where A1, A2, A3, and A4 are the absorbance values ​​of the corresponding reaction tubes at 475nm after the reaction is completed. The tyrosinase inhibition rate of each experimental group is calculated, and the test results are shown in Tables 2 and Figure 1 shown.

[0061] Table 1

[0062] Reaction system A1 A2 A3 A4 Sample to be tested (ul) 0 0 200 200 0.2M, pH=6.80 PBS (ul) 1000 1200 800 1000 500U / mL tyrosinase solution (ul) 200 0 200 0 0.5wt% tyrosine solution (ul) 300 300 300 300

[0063] Table 2

[0064]

[0065]

[0066] From Table 2 and Figure 1 It can be seen that the tyrosinase inhibition rate of the stem cell culture composition provided in Examples 1 to 3 is significantly higher than that of the stem cell culture composition and induction medium provided in Comparative Examples 1 to 5, indicating that the stem cell culture composition provided in the embodiments of the present application has a stronger tyrosinase inhibition rate and can play a whitening role, indicating that the stem cell culture composition induced by the first induction medium containing red safflower flavonoids and the second induction medium containing red safflower flavonoids and ligustrazine has a more significant whitening effect.

[0067] 2. DPPH free radical scavenging test

[0068] Samples to be tested: 8 groups are set, which are the stem cell culture compositions or induction culture media provided in Examples 1 to 3 and Comparative Examples 1 to 5, respectively.

[0069] Test method: Take 200uL of the sample solution to be tested, add 200uL of 0.2mmol / L DPPH anhydrous ethanol solution, mix well, react at room temperature in the dark for 30min, then use an enzyme marker to measure the absorbance of the sample at 517nm, the blank group is 200uL of distilled water and 200uL of DPPH-anhydrous ethanol solution mixture, the control group is 200uL of sample solution and 200uL of anhydrous ethanol mixture, and the DPPH free radical scavenging rate (Q, %) is calculated according to the formula. The results are shown in Table 3 and Figure 2 Q / % = [1-(A1-A2) / A0] × 100%, where A1, A2, and A0 are the absorbance of the sample group, control group, and blank group, respectively.

[0070] Table 3

[0071]

[0072] From Table 3 and Figure 2 It can be seen that the DPPH free radical scavenging rate of the stem cell culture composition provided in Examples 1 to 3 is significantly higher than that of the stem cell culture composition and induction medium provided in Comparative Examples 1 to 5, indicating that the stem cell culture composition provided in the examples of the present application has better antioxidant capacity, and good antioxidant performance can delay skin aging. It indicates that the stem cell culture composition induced by the first induction medium containing flavonoids of red safflower and the second induction medium containing flavonoids of red safflower and ligustrazine has more significant antioxidant and anti-aging effects.

[0073] 3. In vitro proliferation activity of human fibroblasts

[0074] Cells: Human skin fibroblasts HSF, catalog number CB-Y1274, purchased from Shanghai Meiyan Biotechnology Co., Ltd.

[0075] Test samples and grouping: 9 groups were set up, including one negative control group and 8 test groups. The test groups respectively used the stem cell culture composition or induction medium provided in Examples 1 to 3 and Comparative Examples 1 to 5 of the present application, and the DMEM medium was configured to contain 2% v / v of the stem cell culture composition or induction medium and a DMEM medium containing 10% fetal bovine serum by volume. The negative control group used a DMEM medium containing 10% fetal bovine serum by volume.

[0076] Experimental method: Human skin fibroblasts cultured to the 4th to 6th generation logarithmic growth phase were cultured and prepared at a concentration of 1×10 5 The cell suspension was inoculated into a 96-well plate at 200ul / well, with 3 replicates per group, and another group was added with only 200ul DMEM culture medium as a blank control. The 96-well plate was incubated overnight at 37°C and 5% CO2. After the cells adhered, the culture medium was discarded, and the drugs were administered respectively. The culture medium corresponding to the experimental group was added, and the cells were placed in a cell culture incubator for further culture for 24 hours. The culture medium in the wells was discarded, and after washing with PBS, a mixture of 20 μL of MTT reagent (5 mg / mL) and 180 μL of serum-free DMEM was added to each well. After incubation in a cell culture incubator for 4 hours, the culture medium was discarded, 150 μL of DMSO was added, and the cells were oscillated for 10 minutes to dissolve the crystals. The absorbance values ​​of the test group, negative control group and blank control group at a wavelength of 490 nm were detected by an ELISA instrument, and the cell proliferation rate was calculated. The cell proliferation rate is shown in Table 4; cell proliferation rate % = (test group-blank control group) / (negative control group-blank control group) × 100%, wherein the test group: HSF cells + DMEM culture medium + stem cell culture composition; negative control group: HSF cells + DMEM culture medium; blank control group: DMEM culture medium

[0077] Table 4

[0078] Group Cell proliferation rate (%) Example 1 205.12±6.37 Example 2 208.85±7.54 Example 3 203.27±7.82 Comparative Example 1 159.63±4.36 Comparative Example 2 152.54±5.27 Comparative Example 3 156.19±4.38 Comparative Example 4 151.67±4.52 Comparative Example 5 134.54±5.93 Negative control group 100.00

[0079] As can be seen from Table 4, the cell proliferation rate of HSF cells cultured with the stem cell culture composition provided in Examples 1-3 is significantly higher than that in Comparative Examples 1-5, indicating that the stem cell culture composition provided in the examples of the present application can promote cell proliferation, indicating that the stem cell culture composition induced by the first induction medium containing flavonoids of red safflower and the second induction medium containing flavonoids of red safflower and ligustrazine has a more significant effect in promoting the proliferation of HSF cells.

[0080] 4. Content of collagen I secreted by cells

[0081] Cells: Human skin fibroblasts HSF, catalog number CB-Y1274, purchased from Shanghai Meiyan Biotechnology Co., Ltd.

[0082] Test samples and grouping: 9 groups were set up, including one negative control group and 8 test groups. The test groups respectively used the stem cell culture composition or induction medium provided in Examples 1 to 3 and Comparative Examples 1 to 5 of the present application, and the DMEM medium was configured to contain 2% v / v of the stem cell culture composition or induction medium and a DMEM medium containing 10% fetal bovine serum by volume. The negative control group used a DMEM medium containing 10% fetal bovine serum by volume.

[0083] Experimental method: Human skin fibroblasts cultured to the 4th to 6th generation logarithmic growth phase were cultured and prepared at a concentration of 1×10 5 / ml cell suspension, cells were inoculated in 96-well plates at 100ul per well, 3 replicates were set for each group, and cultured overnight in a cell culture incubator at 37°C and 5% CO2; after the cells adhered to the wall, the culture medium was discarded, and the cells were washed with PBS and administered separately, and the culture medium corresponding to the experimental group was added. After incubation in a 37°C incubator for 36 hours, the cell culture supernatant samples were taken, and the collagen content was determined using a human type I collagen (COL-I) ELISA kit (Shanghai Jianglai Biotechnology Co., Ltd.) to evaluate the promotion of type I collagen synthesis in human fibroblasts. The test results of type I collagen content are shown in Table 5.

[0084] Table 5

[0085]

[0086] As can be seen from Table 5, the stem cell culture composition provided in Examples 1-3 has a significantly higher effect on promoting the secretion of type I collagen by HSF cells than that in Comparative Examples 1-5, indicating that the stem cell culture composition provided in the examples of the present application can promote the production of collagen, thereby making the skin smooth and elastic, and has the potential to be used in skin anti-wrinkle repair and anti-aging products, indicating that the stem cell culture composition induced by the first induction medium containing flavonoids from red safflower and the second induction medium containing flavonoids from red safflower and ligustrazine has a good synergistic effect in promoting the secretion of type I collagen by HSF cells.

[0087] The stem cell culture composition provided in the present application can be used in the preparation of skin care products, wherein the skin care products have the effects of anti-aging, skin repair, whitening, etc.

[0088] The above-mentioned embodiments only express the specific implementation of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the present invention. It should be pointed out that for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention.

Claims

1. A placenta / umbilical cord mesenchymal stem cell culture composition, characterized in that: It is the culture supernatant obtained after mesenchymal stem cells are induced and cultured with the first induction medium and the second induction medium; The first induction culture medium comprises red safflower flavonoids, transferrin, bovine serum albumin, linoleic acid, and basal culture medium. The second induction culture medium comprises flavonoids of red sedge, ligustrazine, transferrin, bovine serum albumin, linoleic acid and basal culture medium.

2. A placenta / umbilical cord mesenchymal stem cell culture composition according to claim 1, characterized in that: The first induction culture medium contains 10-30 mg / L flavonoids, 5.0-7.5 ug / L transferrin, 0.4-1.2 ug / L bovine serum albumin, and 2.0-5.5 ug / L linoleic acid.

3. A placenta / umbilical cord mesenchymal stem cell culture composition according to claim 2, characterized in that: The first induction culture medium contains 20 mg / L flavonoids, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid.

4. A placenta / umbilical cord mesenchymal stem cell culture composition according to claim 1, characterized in that: The second induction culture medium contains 10-30 mg / L flavonoids, 60-90 mg / L chuanxiongzine, 5.0-7.5 ug / L transferrin, 0.4-1.2 ug / L bovine serum albumin, and 2.0-5.5 ug / L linoleic acid.

5. A placenta / umbilical cord mesenchymal stem cell culture composition according to claim 4, characterized in that: The second induction medium contains 20 mg / L flavonoids, 75 mg / L chuanxiongzine, 6.0 ug / L transferrin, 0.8 ug / L bovine serum albumin, and 3.5 ug / L linoleic acid.

6. The placenta / umbilical cord mesenchymal stem cell culture composition according to claim 1, characterized in that: The preparation method of red willow flavonoids is as follows: taking dried red willow and crushing it with a grinder to pass it through a 40-mesh sieve to obtain red willow powder; weighing the red willow powder, placing it in a 250mL round-bottom flask, adding 70% volume fraction of ethanol at a solid-liquid ratio of 1g:20ml, and extracting it at 80°C for 2h; filtering while hot, removing ethanol in the filtrate by rotary evaporation, adding redistilled water and an equal amount of petroleum ether, extracting with a separatory funnel, and discarding the petroleum ether layer; extracting the water layer with 20mL of ethyl acetate twice, combining the ethyl acetate layers, and rotary evaporating to dryness to obtain red willow flavonoids.

7. The placenta / umbilical cord mesenchymal stem cell culture composition according to claim 1, characterized in that: The mesenchymal stem cells are derived from placenta or umbilical cord.

8. The placenta / umbilical cord mesenchymal stem cell culture composition according to claim 1, characterized in that: The basic culture medium is DMEM / F12 or α-MEM.

9. The placenta / umbilical cord mesenchymal stem cell culture composition according to claim 1, characterized in that: Its preparation method is: Mesenchymal stem cells were obtained and cultured in a basal medium containing 10% exosome-free fetal bovine serum; The cells were transferred to the first induction medium and cultured for 24 hours. When the cell confluence reached 80%, they were transferred to the second induction medium. After continuing to culture for 36 to 48 hours, the cell culture fluid was collected and centrifuged to obtain a mesenchymal stem cell culture composition, which was stored at 4°C for later use.

10. Use of the placenta / umbilical cord mesenchymal stem cell culture composition according to any one of claims 1 to 9 in the preparation of epidermal whitening, skin repair and anti-aging products.

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