Human platelet lysate as a replacement for fetal bovine serum in human corneal stromal keratocyte and fibroblast culture

• DOI: 10.1111/jcmm.16912
• Publication Date: 2021-09-06T02:41:48Z
• Authors: Nina Seidelmann; Daniela F. Duarte...; Malena Rohde; Sandra Johnen; Sabine Salla; Gary Hin‐Fai Yam; Jodhbir S. Mehta; Peter Walter; Matthias Fuest
• Keywords: Fetal bovine serum; Platelet lysate; Myofibroblast

Abstract The isolation and propagation of primary human corneal stromal keratocytes (CSK) are crucial for cellular research tissue engineering. However, this delicate cell type easily transforms into fibroblasts (SF) scar inducing myofibroblasts (Myo‐SF). Current protocols mainly rely on xenogeneic fetal bovine serum (FBS). Human platelet lysate (hPL) could be a viable, potentially autologous, alternative. We found high survival with both supplements in CSK SF. Cell numbers Ki67+ ratios increased higher fractions hPL FBS detected loss marker expression (Col8A2, ALDH3A1 LUM) increasing the Myo‐SF SMA amounts but decreased incremental substitution types, implying an antifibrotic effect hPL. Immunohistochemistry confirmed RT‐PCR findings. bFGF HGF were only responsible suppressing conversion. Considering all findings, we propose 0.5% as suitable culture, xeno‐free component efficiently preserved characteristics, non‐inferiority terms viability, number proliferation comparison to established protocol.