Culturing bronchial epithelial cell lines involves several steps to establish and maintain the cells in a laboratory setting. Here is a general overview of the process:

1. Cell Line Selection: Choose a bronchial epithelial cell line suitable for your research. Commonly used cell lines include BEAS-2B, Calu-3, and 16HBE14o-.

2. Cell Culture Medium: Prepare a suitable cell culture medium specific to bronchial epithelial cells. This typically involves using a basal medium, such as DMEM or MEM, supplemented with additives like fetal bovine serum (FBS), antibiotics, and growth factors (e.g., epidermal growth factor, insulin).

3. Coating: Coat the culture vessels (e.g., flasks, plates) with extracellular matrix proteins, such as collagen or fibronectin, to enhance cell attachment and growth. Follow the manufacturer's instructions for coating concentrations and incubation times.

4. Thawing Cells: If using frozen cells, thaw the cryovial in a water bath at 37°C, quickly transfer the contents to a tube containing pre-warmed culture medium, and centrifuge to pellet the cells. Gently resuspend the cell pellet in fresh culture medium.

5. Seeding Cells: Add the resuspended cells to the coated culture vessels at the desired seeding density. The seeding density depends on the specific cell line and experimental requirements. Incubate the cells at 37°C in a humidified incubator with 5% CO2.

6. Subculturing: As the cells reach confluence (typically 70-80% confluence), they need to be passaged or subcultured. This involves detaching the cells from the culture vessel using a suitable enzyme (e.g., trypsin-EDTA) and transferring them to new culture vessels at a lower density to allow continued growth.

7. Feeding and Maintenance: Regularly feed the cells by replacing the culture medium every 2-3 days or as needed. Maintain the cells in a favorable environment by ensuring appropriate temperature, humidity, and CO2 levels. Monitor cell growth, morphology, and viability regularly.

8. Quality Control: Perform regular quality control checks, including assessing cell morphology, viability, and contamination. Test the cells for mycoplasma contamination periodically using PCR-based methods or commercial detection kits.

9. Experimental Considerations: Adjust the culture conditions and experimental protocols based on the specific research objectives. For example, you may need to expose the cells to specific stimuli, treat them with drugs, or perform functional assays to assess their responses.

These steps provide a general guideline, and the specific protocols and requirements may vary depending on the chosen cell line, lab protocols, and experimental setups. It is advisable to consult relevant literature, protocols, and experienced researchers in your field to optimize the culture conditions for your specific bronchial epithelial cell line.

Good luck

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Hello Behnoush Nasr Zanjani

You may use the common cell culture media. For instance, 16HBE cell line which is a human bronchial epithelial cell line may be cultured in DMEM supplemented with 10% FBS, 2 mM L-Glutamine, 1% NEAA and 1 mM sodium pyruvate.

Best.

FOR epithelial example

Hi Behnoush Nasr Zanjani

* I have used BEAS-2B cells procured from ATCC. BEAS-2B cells are a human bronchial epithelial cell line that was originally derived from normal bronchial tissue. These cells are commonly used in studies related to respiratory diseases, toxicology, and cellular biology.

The growth media and supplements vary depending on the source of cells. Please find the link for the sources attached.

1. https://www.atcc.org/products/crl-9482

2. https://www.thermofisher.com/in/en/home/technical-resources/cell-lines/b/cell-lines-detail-39.html

3. https://www.sigmaaldrich.com/IN/en/product/sigma/95102433

For detailed information you can also refer to my recent publication.

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Thanks,