Phenotypic transformation of HCC827 cells revealed by evaluation of human platelet lysate as a sustainable fetal bovine serum replacement.

Fetal bovine serum (FBS), a widely used supplement in cell culture, raises ethical and scientific concerns due to its animal origin, batch variability, and limited physiological relevance for human cells. As part of our efforts to adopt more sustainable and human-relevant cell culture conditions, we investigated human platelet lysate (HPL) as an alternative to FBS for culturing non-small cell lung carcinoma (NSCLC) cells. Different compositions of both sera were assessed by ELISA, showing comparable FGF2 content but 2-fold higher amount of TGF-β1 in HPL compared to FBS. Notably, when cultured with HPL, HCC827 cells developed distinct phenotypes, including ring-like f-actin structures, increased spheroid roundness and size. Canonical endothelial-to-mesenchymal transition (EMT) was not detected, supported by Western blot analysis of key markers Vimentin and SNAI1. Instead, a hybrid EMT signaling state based on kinome activity profiling data is visible but cannot fully explain the visible phenotypical changes. Furthermore, kinome activity profiling at different timings revealed significant HPL-dependent changes for HCC827 cells pointing at altered integrin signaling, distinct from those observed in the other NSCLC lines A549 and H1299. Our findings highlight the cell-type-specific effects of HPL compared to FBS and underscore the importance of case-by-case evaluation when considering HPL as an alternative to FBS in cellular models.