Specialized culture medium for rabbit primary adipose derived microvascular endothelial cells

Cell experiment steps:

1、 Cell revival

1. Place 10ml pipette, pipette, 1ml pipette tip, 50ml centrifuge tube, T25 culture bottle, alcohol lamp, waste liquid tank and other items into the ultra clean workbench, irradiate with ultraviolet light for 30 minutes, and then ventilate for 30 minutes;

2. Preheat the culture medium;

3. Disinfect with 75% alcohol and place on an ultra clean workbench;

4. Remove the frozen cells from the liquid nitrogen tank;

5. Place disposable gloves in a 37 ℃ water bath and shake quickly to melt them;

6. Disinfect with 75% alcohol and place on an ultra clean workbench;

7. Transfer 9ml of culture medium into a 50ml centrifuge tube;

8. Use a 1ml pipette tip to aspirate thawed cell suspension into a centrifuge tube;

9.1000r/min, Centrifuge for 5 minutes;

10. Disinfect with 75% alcohol and place on an ultra clean workbench;

11. Discard the supernatant;

12. Take 1ml of culture medium and resuspend the cells. Transfer the cell suspension to a culture bottle, add an appropriate amount of culture medium, gently shake the bottle to evenly distribute the cells, and mark them properly;

13. Observe the density and status of revived cells under a microscope;

14. Put the cells back into the carbon dioxide incubator and let them stand for cultivation.

Product attributes:

The culture medium for rabbit primary adipose microvascular endothelial cells has been carefully optimized by the team. After long-term testing, this product can maintain the excellent growth status of rabbit primary adipose microvascular endothelial cells.

This product already contains various ingredients required for the growth of rabbit primary adipose microvascular endothelial cells, without the need to add any ingredients, and can be directly used for the cultivation of rabbit primary adipose microvascular endothelial cells.

Notes:

For scientific research purposes only.

Some components in the culture system are harmful to human health. Please do not use exposed skin to come into contact with the liquid in the culture system or containers containing residual liquid in the culture system; The concentration and harmfulness of these harmful substances are relatively low. If there is contact, rinse immediately with tap water.

Cell culture steps:

Cell revival:

Remove the frozen cells from the liquid nitrogen tank and quickly thaw them in a 37 ℃ water bath.

After thawing, transfer the cells to a culture dish containing fresh culture medium and gently shake to evenly distribute the cells.

Cultivate in a 37 ℃, 5% CO2 incubator.

Cell fluid exchange:

Remove or discard the old culture medium from the culture dish.

Add PBS buffer and gently rinse the cells to remove debris suspended on the cell surface, repeat several times.

Add new culture medium.

Cell passage:

When the cells grow to 80%~90%, they are passaged.

Remove or discard the old culture medium from the bottle, and rinse with PBS buffer.

Add digestive juice and gently shake to make the digestive juice flow over all cell surfaces.

Place the culture bottle in a 37 ℃ incubator and let it stand for a few minutes to observe changes in the cells. When the intercellular space increases, add culture medium containing serum to terminate digestion.

Blow the cells into a suspension and transfer it to a centrifuge tube for centrifugation.

Discard the supernatant and add an appropriate amount of cell culture medium to resuspend the cells.

Allocate proportionally to new culture dishes and add fresh culture medium.

Mark well and incubate in a 37 ℃, 5% CO2 incubator.

Cell cryopreservation:

Select logarithmic growth phase cells for cryopreservation.

Transfer the cells to a centrifuge tube and centrifuge, discard the supernatant.

Add an appropriate amount of cryoprotectant (such as 90% culture medium+10% DMSO) and gently blow the resuspended cells.

Transfer the cells to a cryopreservation tube, label them, place them in a programmed cooling box, and then freeze them in a -80 ℃ freezer. After a few hours or overnight, transfer to a liquid nitrogen tank for storage.

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