Isolation and Culture of Murine Tongue Epithelial Cells: A Procedure to Isolate and Culture Tongue Epithelial Cells from Mouse Model

The tongue constitutes stratified squamous epithelium and an underlying layer of dense connective tissue termed the lamina propria. The epithelium consists of multi-layered squamous epithelial cells that provide protection against mechanical stress.

To isolate epithelial cells, begin with a freshly harvested mouse tongue. Mince the tongue to obtain smaller tissue fragments. Next, treat the fragments with a customized digestion cocktail containing collagenase, hyaluronidase, and deoxyribonuclease enzymes.

Collagenase and hyaluronidase enzymes digest the extracellular matrix that holds the tissue together, promoting the release of individual cells into the suspension. Deoxyribonuclease degrades any free DNA released from lysed cells.

Add serum-containing buffer to the digested sample to inactivate the protein-hydrolyzing enzymes. Now, filter the sample through cell strainers to remove any undigested tissue fragments and debris and obtain a single-cell suspension of epithelial cells.

Centrifuge to separate the cells. Discard the supernatant containing the digestion mix. Resuspend the cell pellet in fresh media. Finally, transfer the desired volume of this cell suspension into a sterile culture dish. Incubate the culture.

The epithelial cells attach to the bottom surface of the culture dish. Depending on the proliferative capability of individual epithelial cells, they start forming colonies.