Proteomics-supported amniotic membrane extract-supplemented GelMA bioink enables extrusion-based 3D bioprinting of hiPSC-derived corneal stromal constructs
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.
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BibTeX
@article{monla2026proteomicssupported,
title = {{Proteomics-supported amniotic membrane extract-supplemented GelMA bioink enables extrusion-based 3D bioprinting of hiPSC-derived corneal stromal constructs}},
author = {Reem Al Monla and Manon Descieux and Morgane Benard and Melissa Girard and Laura Fichter and Sonia Garcia Lorens and Jerome Vialaret and Christophe Hirtz and Christopher Leon-Valdivieso and Vincent Daien and Marie Rose Rovere and Catherine Moali and Claire Bardet and Frederic Michon},
journal = {Bioprinting},
year = {2026},
month = sep,
doi = {10.1016/j.bprint.2026.e00496},
url = {https://hal.science/hal-05762017v1},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Proteomics-supported amniotic membrane extract-supplemented GelMA bioink enables extrusion-based 3D bioprinting of hiPSC-derived corneal stromal constructs
AU - Reem Al Monla
AU - Manon Descieux
AU - Morgane Benard
AU - Melissa Girard
AU - Laura Fichter
AU - Sonia Garcia Lorens
AU - Jerome Vialaret
AU - Christophe Hirtz
AU - Christopher Leon-Valdivieso
AU - Vincent Daien
AU - Marie Rose Rovere
AU - Catherine Moali
AU - Claire Bardet
AU - Frederic Michon
PY - 2026
DA - 2026/09/24
JO - Bioprinting
DO - 10.1016/j.bprint.2026.e00496
UR - https://hal.science/hal-05762017v1
AB - 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.
SN - 2405-8866
ER -