Graft for descemet's stripping endothelial keratoplasty and mechanical preparation method of graft
A technology of descemet's layer and grafts, applied in medical science, tissue regeneration, prosthesis, etc., can solve the problems of fragility, thin thickness, and difficulty, and achieve the effect of low price, wide source, and large-scale industrialization
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Embodiment 1
[0073] 1. Take the isolated porcine eyeball, and cut out the cornea with 1-2 mm scleral ring on the outer edge. see photos figure 1 (endothelial cell layer placed upwards).
[0074] 2. After completing step 1, perform trypan blue staining on the scleral sulcus (in practice, trypan blue staining can also be performed on the entire cornea obtained in step 1), then rinse with PBS buffer, and then scrape off the trabecular meshwork (pigments attached to the trabecular meshwork). See photos after trypan blue staining figure 2 (endothelial cell layer placed upwards).
[0075] 3. After completing step 2, perform trypan blue staining, tear off part or all of the trabeculae, and expose the edge of the descemet (the exposed edge of the descemet accounts for one-half of the entire edge of the descemet; in actual operation, the exposed The edge of the elastic layer accounts for more than one-fifth of the edge of the entire elastic layer, most preferably one-half). Since the trabecul...
Embodiment 2
[0087] Get the porcine DMEK graft piece that embodiment 1 prepares, detect cell density and live cell ratio.
[0088] The specific method is as follows:
[0089] 1. Take pig DMEK grafts and stain with 0.2% trypan blue staining solution for 2 minutes.
[0090] 2. After completing step 1, wash 2 times with PBS buffer.
[0091] 3. After completing step 2, stain with 1% Alizarin Red S staining solution for 3 minutes.
[0092] 4. After completing step 3, wash 2 times with PBS buffer.
[0093] 5. After step 4 is completed, observe under a microscope and count the cell density (3 high-power fields are randomly selected, and the sum is averaged) and the proportion of viable cells.
[0094] Get the porcine DMEK graft that embodiment 1 prepares, all reach following standard: cell density > 2500 / mm 2 , The proportion of viable cells >90%.
[0095] The graft prepared in Example 1 was used to perform Descemet's endothelial keratoplasty on rhesus monkeys. After continuous observation f...
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