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S. Amano, N. Honda, T. Usui, T. Mimura; An Animal Model of Descemet’S Stripping Automated Endothelial Keratoplasty Using Cultured Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):600.
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Descemet’s stripping automated endothelial keratoplasty (DSAEK) has obtained popularity in the treatment of corneal endothelial dysfunction as an alternative to penetrating keratoplasty. By reconstructing the DSAEK graft using cultured human corneal endothelial cells (HCEC) and corneal stroma, we might solve the shortage of donor corneas. We investigated the feasibility of DSAEK using cultured HCEC in an animal model.
Primary HCEC cultures were established from endothelial cell layer explants. Cell suspension of cultured HCEC at the fifth passage was seeded on a human corneal stroma with a thickness around 150 µm. First, three kinds of insertion technique (Taco folding, Busin glide, and lens glide) were compared. A porcine corneo-scleral graft was settled onto an artificial anterior chamber and DSAEK graft made using cultured HCEC was inserted through a limbal wound of 4.5 mm width using one of the three insertion techniques. After porcine corneo-scleral graft was detached from the artificial anterior chamber, inserted grafts were stained with 1% trypan blue and 1% alizarin red and cellular detachment area was measured using image analysis software (NIH image). Next, feasibility of DSAEK using cultured HCEC was investigated in rabbit eyes. After Descemet’s membrane and corneal endothelial cells in the central area were removed, a DSAEK graft with cultured and DiI-labeled HCEC was attached onto the rear surface of corneal stroma using Busin glide technique and air injection. In a control group, corneal stroma without cultured HCEC was transplanted. Anterior segment was observed with slit-lamp and central corneal thickness was measured with ultrasound pachymeter until one month after the surgery.
Cellular detachment area was larger in Taco-folding group than in Busin glide or lens glide group (p<0.01). One month after the transplantation, the cornea regained clarity in eyes receiving a graft with HCEC whereas the cornea had intense edema in the control eyes. In the eyes that received a graft with HCEC, corneal thickness increased during the first week and gradually decreased. On the other hand, in the control group, corneal thickness was over 1000µm during the follow-up period. Fluorescein microscopic examination on whole mount corneas showed that DiI-positive cells were observed on posterior surface the graft one month after surgery.
DSAEK using cultured HCEC may be an alternative therapy for corneal endothelial dysfunction.
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