Figure 1.
Culture of dermal papilla cells (DPCs) in the DMEM/F12 Medium plus 10% newborn calf serum.
Primary culture of the primary hair follicle-dermal papilla cells (PHF-DPCs) (A, C), and secondary hair follicle-dermal papilla cells (SHF-DPCs) (B, D). The DPCs exhibited a triangular or polygon shape (Figure 1C–F) at primary and subsequent passage. At the second passage both the PHF-DPCs (E) and SHF-DPCs (F) formed cell aggregates during further culture for approximately 20 days. Black arrow in (B) shows the tiny DP isolated from SHF. Scale bars = 100 µm.
Figure 2.
Growth curves of primary hair follicle-dermal papilla cells (PHF-DPCs), secondary hair follicle-dermal papilla cells (SHF-DPCs) and dermal fibroblast cells (DFCs).
The inoculum was 1.0×104 cells/ml. Cell numbers were counted daily and recorded. The population doubling time was approximately 3.6days for PHF-DPC, 3.7days for SHF-DPC and 0.86days for DFC.
Figure 3.
Immunocytochemical analysis of cultured dermal papilla cells (DPCs).
Immunocytochemistry of DPCs using anti-α-SMA antibody (green) was performed (A&B) when cells started to exhibit an aggregative growth behavior, while the other two antibodies were performed on monolayer cultured DPCs. Both primary hair follicle-dermal papilla cells (PHF-DPCs) and secondary hair follicle-dermal papilla cells (SHF-DPCs) were positive for α-SMA (green, A & B), Laminin (red, C & D), and Collagen IV (red, E & F). Nuclei in A–F were marked by DAPI staining (blue). Scale bars = 100 µm.
Figure 4.
Schematic representation of the differentially expressed genes between primary hair follicle-dermal papilla cells (PHF-DPCs) and secondary hair follicle-dermal papilla cells (SHF-DPCs).
Of the 1044 differentially expressed genes, 620 were upregulated (top-left, red) and 424 were downregulated (bottom-right, green) in PHF-DPCs compared with SHF-DPCs.
Figure 5.
Pearson correlation coefficient* of differential expression ratios between RNA-Seq and qRT-PCR.
*Correlation is significant at the 0.01 level.
Table 1.
Validation of ten differentially expressed genes with biological replicates using qRT-PCR.
Table 2.
Upregulated genes in PHF-DPCs that involved in angiogenesis.
Table 3.
Upregulated genes in SHF-DPCs that involved in angiogenesis.
Figure 6.
Differentially expressed genes between primary hair follicle-dermal papilla cells (PHF-DPCs) and secondary hair follicle-dermal papilla cells (SHF-DPCs) involved in extracellular matrix (ECM)-receptor interaction pathway.
The red color labels genes upregulated in the PHF-DPCs compared with the SHF-DPCs. The green color labels genes downregulated in the PHF-DPCs. The yellow color labels genes that some were upregulated and others were downregulated in the PHF-DPCs.
Table 4.
Expression level of differentially expressed genes between PHF-DPCs and SHF-DPCs involved in ECM-receptor interaction pathway.