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Evidence for stem cells in cultures of mouse prostate epithelial cells
Janet A Sawicki1, Craig J Rothman
1Lankenau Institute for Medical Research, Wynnewood, Pennsylvania 19096-3425, USA. sawickij@mlhs.org
Background:
Primary cultures of mouse prostatic epithelial cells were studied to identify cells having stem cell characteristics.
Methods:
Culture conditions supporting the growth of mouse prostatic epithelial cells were identified. Immunostaining identified cells expressing cytokeratin 5 (K5), cytokeratin 8 (K8), and bcl-2. Cells with clonogenic capacity were identified by replating dissociated cells from primary colonies.
Results:
Only 2-4% of prostatic epithelial cells that were plated formed colonies. Most cells cultured for 2-4 days expressed bcl-2, K5, and K8. After 6 days in culture, colonies had two distinct phenotypes, star- or disc-shaped. In colonies of both phenotypes, only a few cells expressed K5 and bcl-2, while all cells expressed K8. Upon replating, two-thirds of star-shaped colonies gave rise to a single colony; of 29 disc-shaped colonies, none gave rise to new colonies.
Conclusions:
Colonies deriving from putative prostatic epithelial stem cells are identified. These cells express K5 and bcl-2 and are clonogenic.