Isolation and Maintenance of Epithelial Stem Cell - Creative Biolabs

Epithelial stem cells are a type of stem cell that resides in the epithelial tissues of the body, which play a critical role in tissue maintenance, repair, and regeneration and are essential for maintaining the structure and function of epithelial tissues. Creative Biolabs, as a leader in the field of stem cell therapies, we are dedicated to developing innovative protocols for the isolation and maintenance of epithelial stem cells. Our state-of-the-art techniques are designed to provide you with the highest-quality stem cells for your research needs.

We will guide you through our comprehensive protocol for the isolation and maintenance of epithelial stem cells. We will cover everything you need to know, from the materials required to the procedure itself, as well as important notes to keep in mind.

Materials Required

• Medium
• Fetal bovine serum
• Penicillin-streptomycin solution
• Trypsin-EDTA solution
• Collagenase and dispase
• Keratinocyte serum-free medium (KSFM)
• Epidermal growth factor

Procedure

Preparation

First, prepare the epithelial stem cell culture dish by coating it with gelatin solution. Incubate the dish at room temperature for 30 minutes, then aspirate the excess gelatin solution.

Isolation

Next, isolate the epithelial stem cells from the donor tissue. Begin by washing the tissue with FBS and penicillin-streptomycin solution. Then, mince the tissue and add collagenase and dispase to the solution. Incubate the mixture at 37°C for 1-2 hours, or until the tissue has dissociated.

Culture

After dissociation, filter the solution and wash the cells. Centrifuge the cells, then aspirate the supernatant and resuspend the cells in KSFM. Once isolate the epithelial stem cells, culture them in KSFM. Incubate the cells at 37°C with 5% CO₂.

Maintenance

Subculture them once they reach 70-80% confluence. Begin by washing the cells, then add trypsin-EDTA solution to the dish. Incubate the dish at 37°C for 2-3 minutes, or until the cells have detached. Neutralize the trypsin-EDTA solution with DMEM containing FBS, then centrifuge the cells and resuspend them in fresh KSFM.

Notes

• It is important to use fresh reagents for each experiment to ensure optimal results.
• The collagenase IV and dispase incubation time may vary depending on the tissue source.
• Be sure to maintain sterility throughout the procedure to prevent contamination.

We hope that this comprehensive protocol for the isolation and maintenance of epithelial stem cells has been helpful. At Creative Biolabs, we are committed to providing you with the most advanced stem cell technologies and protocols available. If you have any questions or comments, please do not hesitate to contact us.

References

1. Yang R, et al. Epidermal stem cells in wound healing and regeneration. Stem cells international, 2020.
2. Lakshmipathy U and Verfaillie C. Stem cell plasticity. Blood reviews, 2005, 19(1): 29-38.

For Research Use Only. Not For Clinical Use.