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Figure 8: Effect of different keratinocyte growth factor (KGF) concentrations on hair follicle stem cells proliferation. Cells were assessed 3 days after
treatment. (A) the KGF 0 µg/L group shows cells have grown well and display a cobblestone-like appearance; (B) the KGF 10 µg/L group shows cell density
has increased. A small number of enlarged cells have appeared; (C) the KGF 25 µg/L group shows differentiated cells with enlarged nuclei and enriched
cytoplasm can be seen; (D) the KGF 50 µg/L group shows the proportion of differentiated cells increased, and the cell density decreased; (E) the KGF 100
µg/L group shows the proportion of differentiated cells increased, cell aging was observed. Magnification, ×100.
β1-integrin expression: HFSCs showed a high rate of β1-integrin
expression, as shown in Figure 5C. Positively stained cells
grew in clusters. Previous studies have confirmed that β1-
integrin is a membrane marker for HFSCs. [5,8]
The influence of LiCl on HFSC proliferation
There were no significant differences between the
proliferation rates of HFSCs treated with lower doses of
LiCl and that of the control group; there was an inverse
correlation between dose and proliferation rate at higher
concentrations of LiCl than 10 mmol/L [Figures 6 and 7]. LiCl
concentrations less than 10 mmol/L had no effect on the
Figure 9: Effect of keratinocyte growth factor (KGF) on hair follicle stem proliferation rate; moreover, treatment with 0-10 mmol/L
cells (HFSCs) proliferation LiCl did not markedly affect cell proliferation, especially for
culture times greater than 72 h (P > 0.05). However, the
were positive for pan-cytokeratin [Figure 5A]. Cytokeratin is proliferation rate decreased at a LiCl concentration of 25
a defining feature of HFSCs. [5,8] mmol/L; there was a clear difference between treated and
control cells at each time point (P < 0.05).
K19-Cy3 expression: isolated keratin 19 (K19)-labeled HFSC
After 24 h of culture, DNA quantitation in the 25 mmol/L
clusters were positive for cytoplasmic K19 [Figure 5B]. K19 group compared with 0 mmol/L group, P < 0.05, as
is a specific cytoplasmic marker for HFSCs. [5,9] compared with other groups, P > 0.05; after 48 h of

Figure 10: Untreated hair follicle stem cells (HFSCs). Cells morphological features of negative controls (HFSCs subculturing without intervention) after 3
(A), 5 (B), 7 (C), and 9 (D) days in culture. HFSC treated with 10 mmol/L LiCl. Cells morphological features shown after 3 (E), 5 (F), 7 (G), and 9 (H) days in
culture. HFSCs treated with 10 µg/L KGF. Cells morphological features shown after 3 (I), 5 (J), 7 (K), and 9 (L) days in culture. Magnification, ×40.
Plast Aesthet Res || Vol 3 || Issue 2 || Feb 29, 2016 43