Isolating corneal endothelial cells

High yield process captures more cells with a lower risk of co-collecting keratocytes

Medical & Biotechnology

Scientists at the Army have developed a device and method for the isolation of corneal endothelial cells (CECs). The patented technology is available via patent license agreement to companies that would make, use, or sell it commercially.

The outline of corneal endothelial cells was highlighted by alizarin red staining. Image: Zhiguo.he

Mammalian CECs are a single layer of cells located on the posterior side of the cornea facing the anterior chamber. CECs play a dual role in eye health by allowing nutrients from the anterior chamber to pass into the cornea and pumping water out of the cornea and into the anterior chamber.

Loss of a critical number of CECs due to disease or injury results in significant inflammation, epithelial bullae, and limbal stem cell deficiency. Together these complications can eventually lead to corneal opacity and total vision loss. As CECs do not regenerate, surgical intervention by corneal transplant is the only available option. However, transplantation is often unavailable due to the limited supply of fresh corneas suitable for transplantation and even in the event of transplantation, tissue rejection is a possibility.

One method for harvesting CECs from donated corneas is Descemet’s stripping. This procedure involves scraping CECs from the underlying basement membrane (Descemet’s Membrane) with a trephine. The procedure can result in the co-isolation of keratocytes located beneath Descemet’s Membrane. Keratocytes are highly proliferative and, CEC isolation by Decemet’s stripping often results in the overgrowth of contaminative keratocytes during cell culture expansion rendering the CECs unusable for transplant.

Army researchers have developed a new apparatus for isolating corneal endothelial cells (CECs). The device uses jets of enzymatic fluid to degrade CDC attachment sites and capture them in a container. The precision of the method and device allows for the capture of only the CECs. This procedure holds promise for effectively treating many more patients through transplant.

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