Transplanted corneal stromal cells in vitreous reproduce extracellular matrix of healing corneal stroma

Abstract

Purpose: To characterize the extracellular matrix (ECM) formed by corneal stromal cells after injection into the vitreous. This will provide a basis for future studies on the function of corneal ECM macromolecules.

Methods: Cell line from rabbit dermal fibroblasts (RAB9) and primary cultures of rabbit corneal stroma fibroblasts (NRCF) were grown to confluence. For each cell type, approximately 1 x 10(6) cells suspended in basal medium were injected into the vitreous of normal rabbits and observed periodically with a slit lamp. After 1,2, and 4 weeks, eyes were processed for transmission electron microscopy (TEM), immunohistochemistry, immunocytochemistry, and in situ hybridization.

Results: All cells showed gradual growth within the vitreous along the needle track. Occurrence of retinal detachment and inflammation was variable. Transmission electron microscopy of NRCF confirmed the deposition of ECM reminiscent of the organization of normal fetal corneal stroma. Similar matrices were produced by RAB9. NRCF deposited collagen fibrils similar in diameter to those seen in normal developing and healing corneal stroma. RAB9 produced collagen fibrils with larger diameters. NRCF-transplanted cells synthesized proteoglycans and collagen immunologically identical to decorin proteins and type VI collagen, indicating that the expression of specific ECM is maintained after transplantation. In addition, in situ hybridization showed that type XII collagen mRNA is synthesized by transplanted NRCF similar to healing corneas.

Conclusions: Corneal stromal cells transplanted into vitreous produce a matrix morphologically and biochemically similar to that in healing corneal stroma.