Method for the Anti-senescence of and/or rejuvenating stem cells
a stem cell and anti-senescence technology, applied in the field of transcinnamaldehyde, can solve the problems of cell cycle arrest, lack of stem cells or progenitor cells, and decrease in stem cell proliferation ra
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example 1
Preparation of Stem Cells
[0071](1) Adipose Stem Cells (ADSCs)
[0072]ADSCs were sub-cultured for 5 to 7 passages in K-SFM (Keratinocyte-Serum Free Medium, purchased from Invitrogen-Gibco) at 37° C. under 5% CO2. The K-SFM contained human recombinant epidermal growth factor (purchased from Gibco), bovine pituitary extract (purchased from Invitrogen-Gibco), 10% of fetal bovine serum (FBS; purchased from Hyclone), 1% of penicillin / streptomycin (P / S; purchased from Biowest), 0.2 mM of L-ascorbic acid (purchased from Sigma), and 2 mM of N-acetyl-L-cysteine (purchased from Sigma).
[0073]Four portions of equal volume of the above culture medium containing the sub-cultured ADSCs were separately handled by adding thereinto H2O2 (purchased from Sigma-Aldrich) at a final concentration of 100 μM and reacting for 2 hours to induce senescence of the ADSCs, replacing the culture medium with a fresh culture medium, adding thereinto TC (trans-cinnamaldehyde, purchased from Sigma-Aldrich) at a final con...
example 2
Effects of H2O2 and TC on the Expression of the Surface Antigens of Stem Cells
[0082]Samples of the “untreated group,”“H2O2 group,” and “H2O2+TC6 group” ADSCs obtained from Example 1 (1) were labelled with the following antibodies: an antibody against human cluster of differentiation 14 (CD 14), an antibody against human cluster of differentiation 29 (CD29), an antibody against human cluster of differentiation 44 (CD44), an antibody against human cluster of differentiation 45 (CD45), and an antibody against human leukocyte antigen-ABC (HLA-ABC) (purchased from Dako Ltd.); an antibody against human cluster of differentiation 34 (CD34), an antibody against human cluster of differentiation 49b (CD49b), an antibody against human cluster of differentiation 73 (CD73), an antibody against human cluster of differentiation 90 (CD90), and an antibody against human leukocyte antigen-DR (HLA-DR) (purchased from Becton Dickinson Ltd.). Then, the expressions of the surface antigens of the cells in...
example 3
Effect of TC on the Differentiation Capacity of Stem Cells
Experiment (1)
Osteogenic Differentiation Assay
[0084]Samples of the “untreated group,”“H2O2 group,” and “H2O2+TC6 group” ADSCs obtained from Example 1 (1) were cultured in DMEM to induce osteogenic differentiation, wherein the DMEM contained 10% of FBS, 20 nM of dexamethasone, 100 Units / mL of penicillin, 100 μg / mL of streptomycin, 2.5 μg / mL of amphotericin, 10 mM of b-glycerophosphate, and 0.05 mM of L-ascorbic acid 2-phosphate. After being cultured for 7 days, the samples were stained with an alkaline phosphatase detection kit (purchased from EMD Millipore, CSCR004) and then photographed by a microscope. The results are shown in FIGS. 2A to 2C, wherein the cells after the osteogenic differentiation were in purple color under the microscope. After being photographed, the absorbance at a wavelength of 450 nm (i.e., OD450) of each sample was measured to figure out the effect of H2O2 and / or TC on the osteogenic differentiation ca...
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