Human gallbladder artery endothelial cells

Human gallbladder artery endothelial cells

Basic cellular properties:

Cell profile: Gallbladder,It is a pear shaped pouch structure located behind the liver below the right rib, which has the function of concentrating and storing bile. The gallbladder wall is composed of three layers: mucosa, muscle layer, and outer membrane. Arteries are composed of intima, media, and adventitia, with the intima mainly composed of endothelial cells.

Culture medium: primary endothelial cell culture system

Fluid change frequency: everyChange the fluid every 2-3 days

Digestive fluid:0.25% pancreatic amylase

Cultivation conditions: Gas phase: Air, 95%; CO2 ，5%

Primary cell culture method:

1. Preparation: Take various disinfected culture products and place them on the purification table. Disinfect with ultraviolet light for 20 minutes. Wash your hands and wipe your elbows with 75% alcohol before starting work.

2. Layout: Ignite the alcohol lamp and install the straw cap.

3. Organize: Place the tissue block in a beaker and rinse 2-3 times with Hanks solution to remove blood stains; If you suspect that the organization may be contaminated, you can first place it in a mixture containing green for 30-60 minutes.

4. Cutting: Use ophthalmic scissors to cut tissue into 2-3 millimeter sized pieces for digestion. Add 30-50 times more liquid than the total amount of tissue blocks, then pour them into a triangular flask together, tie the bottle neck or stopper with a rubber stopper.

5. Digestion: Either use a constant temperature water bath or place in a 37 ℃ incubator for digestion. Shake every 20 minutes during digestion, and it is better to use an electromagnetic constant temperature stirrer for digestion. The digestion time depends on the size of the tissue block and the hardness of the tissue.

6. Separation: When the digestive fluid becomes turbid during the digestion process, a small amount of digestive fluid can be suctioned out with a pipette and observed under a microscope. If the tissue has dispersed into cell clusters or individual cells, digestion should be terminated immediately, and then the tissue blocks that have not been fully digested should be filtered out through a suitable stainless steel sieve. Centrifuge the digestion solution at low speed (500-1000 rpm) for 5 minutes, aspirate the supernatant, and add an appropriate amount of culture medium containing serum.

7. Counting: Use a counting plate to count. If the cell density in the cell suspension is too high, adjust it by adding culture medium and then divide it into culture bottles. For most cells, the pH requirement is in the range of 7.2~7.4, and the culture medium should be slightly red. If the color is yellowish, it indicates that the liquid has become acidic and can be adjusted with NaHCO3.

8. Cultivation: Incubate in a 36.5 ℃ incubator. If CO2 incubator is used for cultivation, loosen the bottle cap by half a turn.

Notes:

1. The cultivation can be stored for 3-6 months under 4 ℃ conditions.

During the cell culture process, please pay attention to maintaining aseptic operation.

During the subculture process, digestion time should not be too long, otherwise it will affect cell adhesion and growth status.

4. It is recommended that customers take several cell photos for each magnification within the first 3 days after receiving the cells, recording the cell status for communication with the technical department; Due to transportation reasons, some sensitive cells may experience instability. Please contact us promptly and provide detailed information about the specific situation of the cells, so that our technical personnel can track and follow up until the problem is resolved.

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