Composition for increasing biological activity of stem cells using mixture 4f
A composition and stem cell technology, applied in the direction of cell culture active agents, animal cells, drug combinations, etc., can solve problems such as difficult to enter the practical stage, aging and function reduction, so as to improve cell survival and implantation survival rate, improve Cell proliferation ability and movement ability, effect of improving blood vessel and tissue regeneration ability
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Embodiment 1
[0124] Example 1. Preparation of Mixture 4F
[0125] Fucoidan (150 nM), tauroursodeoxycholic acid (25 μM), oleuropein (0.5 μM) and vascular endothelial growth factor (2.5 nM) (recombinant human VEGF165) were mixed to prepare mixture 4F. In the examples described later, b-FGF, IGF, EGF, and ascorbic acid added to the existing medium (EndoGo XF medium, Biological Industries, Inc.) were not used, and only 4F was added to the basal medium for experiment.
Embodiment 2
[0126] Example 2. Confirmation of Morphological Characteristics of Vascular Endothelial Progenitor Cells Treated with Mixture 4F
[0127] In order to study the effect of 4F treatment on cell phenotype, the main cell markers were roughly divided into undifferentiated markers (stem cell markers) and endothelial cell lineage markers (Endothelial lineage markers), and analyzed by flow cytometry. Specifically, Mixture 4F (Fucoidan, Tauroursodeoxycholic Acid (TUDCA), Oleuropein, Vascular Endothelial Growth Factor (VEGF)) was mixed with 5% FBS and 1% penicillin / Streptomycin was added together to the basal medium (EBM2, Lonza) for culturing vascular endothelial cells, then treated with various period conditions, and then analyzed for undifferentiated markers using flow cytometry Expression of markers (CD34, CD90, c-kit, CXCR4), endothelial cell lineage markers (Tie2, CD31). To verify the reproducibility of the results, L-EPCs (n=5) were isolated and cultured from cord blood derived ...
Embodiment 3
[0129] Example 3. Confirmation of Cell Migration Ability Treated According to Mixture 4F
[0130] It is known that CXCR4 regulates the migratory ability of stem cells in a manner dependent on the number of days of 4F treatment, and it was confirmed that the expression of CXCR4 is increased. Based on this, SDF-1α, which is a ligand of CXCR4, was analyzed to be necessary for angiogenesis and regeneration. process of cell migration. Specifically, a Transwell migration assay was performed to analyze the ability of cells to migrate due to chemokine stimulation. The 8.0 μm mesh membrane inserts inoculated with cells (5×104 cells) of each experimental group were inserted into the culture plate, and stromal cell-derived factor (SDF-1α) (100 ng / ml) were cultured. After 6 hours, cells detached from the pores of the reticular membrane by stimulation with lower stromal cell-derived factor (SDF-1α) were stained with crystal violet, and the number of migrating cells was counted. SDF-1α ...
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