Culture system for mouse tracheal epithelial cells
Cultured mouse tracheal epithelial are grown in an air/liquid interface culture that allows them to develop differentiated characteristics (i.e., to develop into mucus cells or ciliated cells). This invention may be used for the growth of cells isolated from knockout and transgenic mice. The primary culture cells of the culture may be ciliated or non-ciliated differentiated cells.
1. A method for producing a culture of differentiated, mouse airway epithelial cells comprising at least 5% mucus cells comprising:
obtaining tracheal epithelial cells from a mouse; and then
maintaining the cells at an air-liquid interface to produce a culture of mouse airway epithelial cells comprising at least 5% mucus cells, whereby the cells remain differentiated.
2. The method according to claim 1 , whereby the maintenance step comprises:
growing the tracheal epithelial cells on a solid support, wherein a first side of the support is in contact with a basolateral medium, and a second side of the support is in contact with an apical medium;
removing the apical medium to create the air-liquid interface; and
continuing to grow the epithelial cells at the air-liquid interface.
3. The method according to claim 2 , wherein the solid support is a semi-permeable membrane.
4. The method according to claim 2 , wherein the basolateral medium comprises serum.
5. The method according to claim 2 , wherein the basolateral medium comprises 5% to 10% serum.
6. The method according to claim 2 , wherein the basolateral medium comprises serum until the air-liquid interface is created.
7. The method according to claim 1 , wherein the mouse is a transgenic mouse.
8. The method according to claim 1 , wherein the mouse is a knockout mouse.
9. The method according to claim 1 , wherein the tracheal epithelial cells produce mucin.
10. The method according to claim 1 , wherein the culture further comprises ciliated cells.
11. The method according to claim 1 , wherein the culture further comprises goblet cells.
12. The method according to claim 1 , wherein the tracheal epithelial cells proliferate in response to an inflammatory mediator.
13. The method according to claim 12 , wherein said inflammatory mediator is IL-13.