3D engineered corneal stroma-derived cells on a laser-assisted bioprinted matrix

Abstract

Purpose : To print and characterize human corneal stroma-derived cells (CSCs) in collagen-containing bioink using a laser-assisted 3D printing method.

Methods : CSCs were isolated from the central corneal-stroma using the explant method, and expanded in standard culturing medium, supplemented with 10% FBS. The cells were then printed in collagen containing bioink, positioned on the top of 1 mm thick stabilizing Matriderm® sheets, then cultured further and processed for immunocytochemistry and transmission electron microscopy.

Results : The CSCs formed dense, multi-layered structures on the Matriderm® sheets with viable cells. Proliferation was demonstrated by Ki-67 positivity in the constructs, comparable to the 2D controls. The printed cells mostly resided on the upper layer during the follow-up period. Expression of mesenchymal stem cell (MSC)-associated and other putative stemness markers was detected. De novo deposition of collagen I could be detected, forming the basal layers of the construct, while expression of keratocyte markers was negative.

Conclusions : CSC-containing constructs have been successfully generated using a laser-assisted bioprinting method. The printed cells showed high viability and proliferative capacity. The printed CSCs demonstrated expression of MSC markers comparable to the 2D cultured cells. Constructs presented here may provide a method for standard production of biocompatible materials using cells, with possible therapeutic potential, as an alternative to corneal allotransplantation in the future. Ample amount of studies has shown the beneficial effects of CSCs in assisting wound healing affecting the corneal stroma.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.