Evaluating the Characteristics of Human Hair Follicle Dermal Papilla Cells in Aggregation Culture Procedure
Vinh Xuan Le, Thuy Thi-Thanh Dao, Nhu Thi-Ngoc Nguyen, Loan Thi-Tung DangBackground:
Human hair follicle dermal papilla cells (hHF-DPCs) play a critical role in hair follicle regeneration. Under conventional two-dimensional (2D) culture conditions, DPCs progressively lose the expression of key biological markers, suggesting that three-dimensional (3D) culture may better preserve their characteristic properties.
Methods:
In this study, hHF-DPCs were seeded into 0.2 mL polypropylene microtubes at three cell densities – 5 × 10
4
, 10
5
, and 2 × 10
5
cells/mL – to evaluate forced aggregation as a 3D culture approach. Aggregate formation and diameter were monitored over time. Viability was assessed by acridine orange/propidium iodide (AO/PI) staining and Trypan blue exclusion, while gene expression of
Results:
Results demonstrated that hHF-DPC aggregates formed within 24 h across all densities. At 5 × 10
4
cells/mL, spheroids formed but disaggregated by day 3, whereas higher densities (10
5
and 2 × 10
5
cells/mL) produced compact, architecturally stable spheroids maintained for 7–10 days. Necrotic cores were detected from day 3 at the two higher densities, accompanied by a reduction in viable cell number. Compared with 2D culture, 3D spheroids exhibited significant upregulation of
Conclusions:
Microtube-based forced aggregation successfully generated hHF-DPC spheroids at densities of 10 5 and 2 × 10 5 cells/mL within 24 h, with stable morphology maintained for up to 10 days and enhanced expression of DPC-specific markers compared to conventional 2D culture.