Application of umbilical cord stem cells in preparation of anti-aging medicines and related beauty products
A technology of umbilical cord stem cells and beauty products, applied in the direction of drug combinations, animal cells, vertebrate cells, etc., can solve problems such as toxic and side effects, achieve the effects of inhibiting aging, good anti-aging activity, and improving skin moisture content
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Embodiment 1
[0031] Example 1 Screening of drugs that specifically inhibit the activity of serine threonine protein kinase mTOR
[0032] According to the method in the prior art "Establishment of the Molecular Level Screening Model of Anti-tumor Drugs Targeting Protein Kinase IKKβ and mTOR", the molecular level drug screening model targeting mTOR was constructed, specifically, it was prepared by using the insect cell baculovirus expression system The mTOR truncated mutant mTOR-FKD with biological activity capable of phosphorylating substrates was produced. The contents of each component in the model were mTOR-FKD 4ng, ATP0.6μM, substrate polypeptide 2μM, and the mTOR inhibitor positive compound PI-103 was used as the As a positive screening control, using the random peptide library constructed by the inventor as the screening target, a total of 7 polypeptides with inhibitory activity were screened, among which TOR-4 and TOR-6 polypeptides had the strongest inhibitory effect, with an IC50 va...
Embodiment 3
[0036] Example 3 Effect of TOR-4 polypeptide on umbilical cord blood stem cells
[0037] Take well-grown cells, trypsinize and stain the cells to confirm that the viable cell count is above 95%, and adjust the cell concentration to 1×10 with DMEM culture fluid containing 10% FBS and VEGF (10ng / m1). 5 / ml, respectively added to 96-well cell culture plate, 0.1mL per well, placed in 5CO 2 1. Cultivate in a 37°C incubator for 24 hours, and add 0.1ml of the above-mentioned culture solution containing TOR-4 polypeptide after the cells adhere to the wall. The experiment was divided into 6 groups to add samples, the concentrations were 0.1, 1, 10, 100, 1000ng / ml, culture medium was added to the control group, and the positive control was rapamycin, the concentrations were 0.1, 1, 10, 100, 1000ng / ml , each group is set up 6 duplicate wells, after cultivating 96h, conventional operation, according to measuring wavelength 492nm application microplate reader, measure the absorbance (A) v...
Embodiment 4
[0041] Example 4 Effect of TOR-4 polypeptide on apoptosis of umbilical cord blood stem cells
[0042] After inoculation of single-cell suspension of umbilical cord blood stem cells, culture in serum-free DMEM medium for 24 hours to synchronize the cell cycle. The cultured cells were divided into control group and experimental group. The control group was added with culture medium, and the experimental group was added with culture medium containing different concentrations of TOR-4 polypeptide. solution (10, 100, 1000ng / m1), continue to culture for 72h, digest and collect the cells to be processed and detected by flow cytometry, the experiment was repeated three times. The results are shown in Table 2.
[0043] Table 2 Effects of different concentrations of TOR-4 polypeptides on stem cell apoptosis
[0044] group Apoptosis rate (%) control 6.21±0.32 TOR-4 polypeptide 10ng / mL 0.65±0.07* TOR-4 polypeptide 100ng / mL 0.53±0.05* TOR-4 polypeptide 1...
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