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Isolation of Human Oral Keratinocyte Progenitor/Stem Cells

Department of Oral and Maxillofacial Surgery, University of Michigan Health System, B1-208 TC, Box 0018, 1500 East Medical Center Drive, Ann Arbor, MI 48109-0018, USA.

^* corresponding author, sefein{at}med.umich.edu

Progenitor/stem cell populations of epithelium are known to reside in the small-sized cell population. Our objective was to physically isolate and characterize an oral keratinocyte-enriched population of small-sized progenitor/stem cells. Primary human oral mucosal keratinocytes cultured in a chemically defined serum-free culture system, devoid of animal-derived feeder cells, were sorted by relative cell size and characterized by immunolabeling for ß1 integrin, nuclear transcription factor, peroxisome proliferator-activated receptor-gamma, and cell-cycle analysis. Sorted cells were distinguished as progenitor/stem cells by functional assays and their ability to regenerate an oral mucosal graft. Small-sized cells demonstrated the lowest expression of peroxisome proliferator-activated receptor-gamma, the highest colony-forming efficiency, a longer long-term proliferative potential, an enriched quiescent cell population, and the ability to regenerate an oral mucosal graft, implying that the small-sized cultured oral keratinocytes contained an enriched population of progenitor/stem cells.

KEY WORDS: oral mucosa • keratinocyte • progenitor cell • stem cell • isolation