What are epithelial cells

There are many cells in the human body, and different cells have different functions. Therefore, we need to have a comprehensive understanding of cells. When there is a problem with human cells, we know what kind of damage it will cause to human health. The preferences of different parts of the human body are different, and the number is also different. So what are epithelial cells? Many people are not very clear about this issue.

Many people do not know what low epithelial cells are. You can get detailed consultation on such issues so that when culturing epithelial cells, you will know what method is the best to choose without harming your health.

What are epithelial cells?

Epithelial cell culture

1) Epidermal cell culture

1. Material collection: Take small pieces of surgical skin grafts or residual skin from surgery, preferably with a thin keratinized layer, and the skin of premature aborted babies is even better, and cut into small pieces of 0.5 to 1 square centimeter.

2. EDTA treatment: first place in 0.02% EDTA at room temperature for 5 minutes.

3. Cold digestion: Replace with 0.25% trypsin and incubate at 4°C overnight.

4. Separation: Remove the skin and use forceps or tweezers to separate the epidermis and dermis.

5. Warm digestion: Remove the epidermis and process it separately. Cut it into smaller pieces with scissors, place it in new 0.25% trypsin, and digest it at 37℃ for another 30 to 60 minutes.

6. Use a pipette to blow up and down gently to make a cell suspension.

7. Culture medium: After filtering through an 80-mesh stainless steel mesh, centrifuge at low speed, aspirate the supernatant, directly add Eagle solution and 20% calf serum to make a cell suspension, inoculate into a dish, and culture in a CO2 incubator.

2) Breast tissue culture

Direct culture method: (suitable for culturing soft tissue with little fiber)

1. In a container containing a small amount of culture medium or Hanks' solution, use a sharp blade to repeatedly cut the tissue into pieces.

2. Inject the tissue fragments and liquid into a centrifuge tube, add a small amount of culture medium, blow with a pipette for a while, and place in a test tube rack for 3 to 5 minutes. Aspirating the supernatant fluid can eliminate non-mammary cell parts. Repeat 2 to 3 times.

3. After the last treatment, add more culture medium to the sedimentation tube and blow lightly with a pipette to resuspend the sediment. Without waiting for the cell clumps to drop, filter through 3 to 4 layers of sterile gauze into another tube.

4. Adjust the appropriate density and inoculate into culture bottles for cultivation.

Collagenase digestion method: (suitable for processing harder tissues containing more fibers)

The process is the same as culturing other tissues.

3) Gastric epithelial cell culture

1. Sampling: Take a small amount of mucosa from the non-lesioned area at the distal end of the gastric specimen removed during gastric ulcer or gastric cancer surgery.

2. Cleaning: After rinsing with a solution containing gentamicin (400 μg/ml) and amphotericin (2 μg/ml in Hanks), peel off the lower mucosa with a blunt instrument and cut into 1 mm3 sizes.

3. Digestion: Digest in type I collagenase and hyaluronidase at 37°C for 80 minutes.

4. Centrifugation: Collect the cell suspension, centrifuge at 800 rpm, and rinse twice with Hanks solution.

5. Inoculation: After the last centrifugation, add complete culture medium containing 1% to 2% fetal bovine serum and inoculate into culture plates with different numbers of wells. The inoculation amount depends on the purpose of the experiment.

4) Hepatocyte culture

Primary tissue block culture: Take a fresh liver, first remove the fibrous components such as the membrane and blood vessels, use a knife or scissors to cut the liver into small pieces of about 1mm3, and use the wall-attached culture method.

5) Endothelial cell culture

1. Take the fresh umbilical cord after delivery. If not cultured immediately, it can be stored at 4°C, but not for more than 12 hours. Aseptically cut into 10-15 cm long sections. Others, such as large blood vessels of embryos and young animals, can also be used for culture.

2. First use a three-way syringe to absorb warm PBS solution and inject it into the umbilical cord vein to wash away residual blood. The injection port should be tied with a string to prevent liquid reflux.

3. Clamp one end of the umbilical cord with a vascular clamp, and slowly inject collagenase with a final concentration of 0.1% into the umbilical vein from the other end. After liquid appears at the end, ligate it to fill the blood vessels. The injection port should also be ligated to prevent liquid reflux. Digest for 3 to 10 minutes.

4. Aspirate the digestion fluid containing endothelial cells and inject it into a centrifuge tube. To obtain more cells, inject warm PBS to rinse 2 to 3 times to thoroughly remove the remaining cells and inject them into the centrifuge tube for centrifugation.

5. Aspirate the supernatant, add 1640 culture medium to make a cell suspension, inoculate into flasks and culture. If all goes well, the cells can grow into a monolayer within 2 to 3 days.

Through the above introduction, we have some introduction to what epithelial cells are and what are the methods of culturing epithelial cells. These are all explained in detail. However, when culturing them, you cannot force it. You must choose according to your own needs. This will be of great help to all aspects of the human body.