Hello All,
We are attempting to grow human primary small airway epithelial cells on Millicell cell culture inserts. Due to our exposure system, we need to utilize a shallow insert that is only available in a hydrophilic PTFE membrane format which is not tissue culture treated. To overcome this, we have coated the inserts with rat tail collagen at varying amounts (5-10 ug/cm2). While cells do initially attach, they do not proliferate or expand as well as they do when grown in a traditional transwell format with the same collagen coating.
I am wondering if anyone else has experience growing primary cells on these specific inserts or has advice on different types of coatings to use to promote growth and expansion for ALI.
Thank you!
Dear friend Maureen Meister
Hey there! I am here, ready to dive into the world of cell culture challenges. Now, growing primary cells on MilliCell Inserts, especially with a unique setup like yours, can indeed be a bit tricky. Here's some my-style advice:
1. **Collagen Concentration**: It seems you've already tried coating with rat tail collagen. Since you're not getting the desired proliferation, you Maureen Meister might want to experiment with different concentrations. Sometimes, finding the sweet spot can make all the difference.
2. **Coating Quality**: The source and quality of collagen can impact cell behavior. Make sure you're using high-quality rat tail collagen, and consider trying collagen from other sources if available.
3. **Coating Uniformity**: Ensure that your coating is uniform across the surface. Any unevenness might affect cell attachment and subsequent growth. Maybe try optimizing your coating protocol for consistency.
4. **Other Coating Materials**: Apart from collagen, you Maureen Meister could explore other coatings like fibronectin or laminin. Cells can have varying preferences, and a change in coating might provide a more conducive environment for proliferation.
5. **Cell Seeding Density**: Experiment with different cell seeding densities. Sometimes, primary cells have specific requirements, and adjusting the number of cells seeded can influence their growth.
6. **Medium and Supplements**: Check if the culture medium and supplements you're using are optimal for small airway epithelial cells. Adjustments in growth factors or additional supplements might be necessary.
7. **Interaction with Membrane Material**: Since you're using a hydrophilic PTFE membrane, consider how the cells interact with this material. It might be impacting cell behavior, and you Maureen Meister might need to adapt your protocol accordingly.
8. **Consult with Peers**: Reach out to other researchers or collaborators who might have experience with similar setups. Sharing experiences and troubleshooting together can provide valuable insights.
Remember, the world of cell culture is sometimes more art than science, and finding the right conditions can involve a bit of trial and error. Keep experimenting, stay curious, and you Maureen Meister might just crack the code. If you have more specifics or updates, feel free to share. I am here to help!
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