Human umbilical vein endothelial cells|378266 |BNCC

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HUVEC

Culture medium	BNCC358173
Description	Endothelial Cell-Specific Medium
Composition	500mlECM+25mlFBS+5mlECGS+5mlP/S
Growth conditions	37°C; 5% CO₂ + 95% air
Subculture procedure	Resuscitation steps: ① Remove the Frozen vial from liquid nitrogen or -80 ℃ refrigerator and place it in PE gloves, quickly immerse it in a 37 ℃ water bath, shake the Frozen vial to accelerate dissolution, and dissolve it completely within 1 minute; ② Add the dissolved cell solution into a centrifuge tube containing 9mL of complete culture medium on a super clean bench, centrifuge at 1000-1200rpm for 5 minutes, discard the supernatant, and resuspend the cells in 1-2mL of complete culture medium. ③ Add the cell suspension into a T25 bottle containing 5-6mL of complete culture medium and incubate in a culture incubator. Cell passage: 1. Discard the culture medium in the bottle and rinse twice with PBS. 2. Add 2.5mL trypsin (T/E solution) and gently shake the flask to ensure complete coverage of the cells with the T/E solution. Monitor changes in cell morphology with a microscope. Once the cells begin to round, transfer the T/E solution from the flask to a 15 ml centrifuge tube (a small portion of cells may separate) and continue incubating the flask at 37 ° C for 1 minute (when there is no solution in the flask). At the end of incubation, gently tap one side of the flask to remove the cells from the surface. Inspect under a microscope to ensure that all cells have shed. 5. Add 5 ml TNS solution to the flask and transfer the separated cells to a 15 ml centrifuge tube. Rinse the flask with 5 ml of TNS and collect any remaining cells. 6. Observe under a microscope whether the flask has successfully harvested cells and observe the number of cells left behind; It should be less than 5%. 7. Centrifuge a 15 ml centrifuge tube at 1000 rpm for 5 minutes. Gently resuspend the cells in the culture medium. 8. Pay attention to changes in the pH value of the culture medium and cell density, change the medium regularly (2-3 times a week), and repeat the passage operation or cryopreservation when the cell density reaches 80% -90%.
morphology	Endothelial cells, Spindle-shaped ，Monolayer adherent growth
Sharing mode	Public welfare sharing