Work

Proteomics-supported amniotic membrane extract-supplemented GelMA bioink enables extrusion-based 3D bioprinting of hiPSC-derived corneal stromal constructs

Year: 2026

Type: article

Abstract: Corneal stromal bioprinting requires bioinks that combine optical transparency, structural stability, printability, and a biologically instructive microenvironment capable of maintaining keratocyte quiescence. Here, we report the development of GAME, a composite bioink integrating gelatin methacryloyl (GelMA) with murine amniotic membrane extract (AME), and comprehensively characterize its biochemical composition, physicochemical properties, bioprintability, and ability to support human induced pluripotent stem cell-derived corneal stromal keratocyte-like cells (hiPSC-CSKs). Quantitative proteomic analysis demonstrated that AME possesses a conserved proteome enriched in collagens, small leucine-rich proteoglycans, fibronectin, nidogens, and growth factor-associated regulatory proteins, with compositional consistency across independently prepared batches. Incorporation of AME into GelMA generated a transparent hydrogel with an interconnected porous microarchitecture, sustained structural stability, and elastic-dominant viscoelastic behavior within the range reported to support corneal stromal cell function. The GAME bioink also tolerated surgical suturing and enabled reproducible extrusion bioprinting of corneal-scale constructs with high printability and geometric fidelity following photocrosslinking. Bioprinted hiPSC-CSKs embedded in GAME exhibited high viability throughout 21 days of culture while maintaining expression of the keratocyte-associated markers keratocan, ALDH3A1, and CD34, with minimal α-smooth muscle actin (α-SMA) expression, indicating preservation of a quiescent, non-fibrotic stromal phenotype after bioprinting. These findings establish GAME as a compositionally defined, mechanically robust, and cytocompatible bioink for corneal stromal tissue engineering. More broadly, this work demonstrates that proteomic characterization of biologically derived supplements provides a rational framework for the development of reproducible and biologically instructive bioinks for corneal tissue engineering.

Source: Bioprinting

DOI: 10.1016/j.bprint.2026.e00496