Database-guided identification of high-performance cell line for robust human cell-derived ECM hydrogel fabrication

Extracellular matrix (ECM) hydrogels are essential for recapitulating native microenvironments in fundamental biomedical research, yet conventional animal tissue-derived products face challenges of cross-species variability or donor-related inconsistency. Human cell-derived matrix (hCDM) fabricated via cell sheet technology offers a promising alternative; however, its efficient fabrication remains constrained by the limited expansion capacity and inconsistent ECM deposition behavior of commonly used primary cells. To facilitate rational cell-source selection, we established a database-guided screening strategy integrating extracellular matrix-related expression profiles, proliferative characteristics, and commercial accessibility. Using primary human dermal fibroblasts (HDFs) as a functional reference, Hs 578 T cells emerged as a top-ranked candidate exhibiting strong ECM deposition potential together with robust proliferative capacity. In vitro validation confirmed that Hs 578 T cells exhibited ECM deposition capacity significantly exceeding that of HDFs. Under optimized serum-reduced conditions, Hs 578 T cells formed cohesive, protein-rich cell sheets that were successfully processed into structurally stable hCDM hydrogels retaining abundant collagens and other critical ECM components. Functional assessment demonstrated that the resulting hCDM hydrogel supports endothelial cell culture and three-dimensional vascular network formation at levels comparable to collagen type I hydrogel. These findings establish a database-guided workflow for rational seed cell selection, providing a strategy that bridges cell sheet cultivation with the efficient fabrication of human ECM biomaterials. STATEMENT OF SIGNIFICANCE: Developing human ECM biomaterials via cell sheet technology is frequently constrained by the inherent variability and limited expansion of primary seed cells. This study introduces a database-guided screening strategy integrating transcriptomic profiles, growth kinetics, and commercial availability to systematically identify high-performance human cell lines for matrix fabrication. By targeting cells with superior biosynthetic and proliferative traits, we demonstrate that a representative cell line, Hs 578 T, can produce ECM-rich substrates with enhanced efficiency and consistency compared to conventional fibroblasts. Under optimized conditions, Hs 578 T cells formed protein-rich cell sheets processed into ECM hydrogels retaining native complexity and pro-vascular bioactivity. This workflow enables rational seed cell prioritization, bridging cell sheet technology with the efficient fabrication of bioactive human ECM materials.