Ann Dermatol.  2009 Nov;21(4):364-368. 10.5021/ad.2009.21.4.364.

Reduced Mitochondrial Properties in Putative Progenitor/Stem Cells of Human Keratinocytes

• Chang SE
• Pak YK
• Lee HW
• Choi JH
• Jeong EJ
• Choi SH
• Chang HW
• Chung YS
• Kim SY

• KMID: 2156498
• DOI: http://doi.org/10.5021/ad.2009.21.4.364

Abstract

BACKGROUND
The characterization of progenitor/keratinocyte stem cells (KSC) remains an unachieved goal. A previous study showed that rapid adhering cells to collagen IV had the characteristics of putative progenitor/KSCs.
OBJECTIVE
The purpose of this study was to investigate the genetic expression of rapid adhering cells compared to non adhering cells to determine the characteristic of KSCs.
METHODS
We isolated rapid adhering cells representative of KSCs from non adhering cells representative of transient amplifying cells. In addition, we differentiated cells from human tonsilar keratinocytes utilizing the adhering capability of the KSCs to collagen IV. Annealing control primer based differentially displayed polymerase chain reaction (PCR) was performed as well as Western blot analysis.
RESULTS
The levels of mitochondria-related gene expression were low in the rapid adhering cells compared to the non adhering cells. Mitochondrial complex I, COX IV, peroxiredoxins (I, II and IV) and mitochondrial membrane potential were all low in the rapid adhering cells compared to the non adhering cells.
CONCLUSION
Using an adhesion method on human collagen IV-coated plates, our results suggest that reduced mitochondrial function may be an important characteristic of KSCs.

Keyword

Adhesion; Keratinocyte progenitor/stem cell; Mitochondria

MeSH Terms

Blotting, Western
Collagen
Gene Expression
Humans
Keratinocytes
Membrane Potential, Mitochondrial
Mitochondria
Peroxiredoxins
Polymerase Chain Reaction
Stem Cells
Collagen
Peroxiredoxins

Figure

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  Fig. 1 Mitochondrial function in the R.A. and N.A. cells. Primary cultures of oral tonsilar mucosal keratinocytes were established and separated by their different adhesion to collagen type IV. The expression of mitochondrial proteins in R.A. and N.A. cells was confirmed by Western blot analysis. (A) The expression of mitochondrial complexes in R.A. and N.A. cells. (B) The expression of peroxiredoxin 1~6 in R.A. and N.A. cells.

• 

  Fig. 2 TMRE flow cytometry and confocal microscopy for mitochondria. (A) R.A. (dotted line) and N.A. (solid line) cells were stained with TMRE and analyzed by FACS. The abscissa shows the intensity of the fluorescence, the ordinate the relative number of cells. (B) R.A. and N.A. cells were treated with MitoTracker Orange CMTMRos to stain the mitochondria and then studies under a confocal microscope.

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