Lentiviral hTERT Overexpression Extends the Lifespan of Primary Human Epidermal Melanocytes and Reshapes Transcriptional Programs
Year: 2026
Type: article
Abstract: Primary human epidermal melanocytes are valuable models for pigmentation research, but maintaining prolonged expansion together with stable melanocytic characteristics can be challenging. Here, primary melanocytes were transduced with an hTERT-overexpression lentiviral vector to generate a mixed population (MIX) and three single-cell-derived clones (SC1, SC3, and SC4). RNA sequencing was performed on passage-6 primary melanocytes, MIX cells, SC clones, and three melanoma cell lines (A375, WM-115, and SK-MEL-1). hTERT overexpression extended melanocyte culture lifespan by more than 10 passages. TERT expression increased in MIX cells and was highest in SC clones. Principal component and gene-expression analyses showed that MIX and SC cells remained globally closer to primary melanocytes than to melanoma cell lines, without an evident melanoma-like transcriptomic shift. However, lifespan extension and clonal selection were accompanied by marked morphological changes and broad transcriptional remodeling involving cell-cycle regulation, telomere-associated processes, antiviral responses, extracellular matrix organization, adhesion, dendrite-related programs, and pigmentation. Melanocyte differentiation and pigmentation programs were generally reduced, particularly in clonal cultures. These findings support hTERT-extended melanocytes as expandable in vitro models, while highlighting that hTERT expression and single-cell cloning can induce functional model drift.
Source: Biomolecules
DOI: 10.3390/biom16091239
Authors: Qiaohua Li, Shaoxuan Liu, Jingxing Gao, Jie Li, Hong Shi
Institution: Kunming University of Science and Technology
Language: English
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,258
OpenAlex ID: W7204467463