LAUSR

Current knowledge gaps on tendon tissue healing can partly be ascribed to the limited availability of physiologically relevant culture models. An unnatural extracellular matrix, high serum levels and random cell morphology in vitro mimic strong vascularization and lost cell elongation in pathology, and discord with a healthy, in vivo cell microenvironment. The thereby induced phenotypic drift in tendon‐derived cells (TDCs) compromises the validity of the research model. Therefore, this research quantified the extracellular matrix (ECM)‐, serum‐, and cell morphology‐guided phenotypic changes in tendon cells of whole tendon fascicle explants with intact ECM and TDCs cultured in a controlled microenvironmental niche. Explanted murine tail tendon fascicles were cultured in serum‐rich or serum‐free medium and phenotype was assessed using transcriptome analysis. Next, phenotypic marker gene expression was measured in in vitro expanded murine tail TDCs upon culture in serum‐rich or serum‐free medium on aligned or random collagen I patterns. Freshly isolated fascicles or TDCs served as native controls. In both systems, the majority of tendon‐specific genes were similarly attenuated in serum‐rich culture. Strikingly, 1‐week serum‐deprived culture—independent of cell morphology—converged TDC gene expression toward native levels. This study reveals a dynamic serum‐responsive tendon cell phenotype. Extracting fascicles or TDCs from their native environment causes large changes in cellular phenotype, which can be limited and even reversed by serum deprivation. We conclude that serum‐derived factors override matrix‐integrity and cell morphology cues and that serum‐deprivation stimulates a more physiological microenvironment for in vitro studies.