A method for long-term culture of animal cornea to evaluate eye irritation damage and repair
A stimulating, corneal technology, applied in the direction of biochemical equipment and methods, animal cells, vertebrate cells, etc.
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Embodiment 1
[0111] Embodiment 1 SLS is to the damage of porcine cornea and evaluation of natural repairing effect
[0112] 1. Preparation of isolated porcine cornea
[0113] 1.1 Porcine cornea preparation: The eyeballs were removed from the orbit within 2 hours after the pig was slaughtered, and the cornea was avoided during the separation process. The isolated porcine eyeballs were transported to the laboratory completely submerged in ice-cold HBSS balanced salt solution containing amphotericin B. The excision of pig eye cornea should be carried out in the sterile area of the laboratory. Before cutting the cornea, the integrity of the cornea should be checked visually for scratches, pigmentation, turbidity and grape sores; if the eyeballs with the above defects should be discarded. Immerse pig eyeballs in 1% povidone-iodine solution for 2 minutes, wash them with sterile PBS, immerse them in PBS containing 0.1% gentamicin for 15 minutes, and perform corneal resection after disinfection...
Embodiment 2
[0139] Example 2 Bovine corneal mechanical damage and natural repair evaluation
[0140] 1. Ex vivo Cornea Preparation and Culture
[0141] 1.2 Bovine cornea preparation: The eyeballs were removed from the orbit within 5 hours after slaughter, and the cornea was avoided during the separation process. The isolated bovine eyeballs were transported to the laboratory completely submerged in HBSS containing penicillin / streptomycin at 4°C. The condition of the bull's eye was inspected prior to harvesting the cornea and discarded if any significant vascular changes, pigmentation, cloudiness, grape sores, or mechanical scratches were observed. The detached cornea was resected from the intact eyeball, leaving 2-4 mm of intact annular sclera at the edge of the cornea. The isolated cornea was continuously washed 3-5 times with 5-7ml sterile HBSS.
[0142] 1.2 Gel filling: Take the cleaned bovine cornea and suspend it in a 6-well plate with the corneal epithelium facing down. Add drop...
Embodiment 3
[0161] Example 3 The repairing effect of recombinant human fibroblast growth factor on pig cornea after injury
[0162] 1. Preparation and Culture of Isolated Porcine Cornea
[0163]1.1 Porcine cornea preparation: The eyeballs were removed from the orbit within 24 hours after the pig was slaughtered, and the cornea was avoided during the separation process. The isolated porcine eyeballs were transported to the laboratory completely submerged in ice-cold HBSS balanced salt solution containing amphotericin B. The excision of pig eye cornea should be carried out in the sterile field of the laboratory, and the complete and defect-free cornea should be excised for visual inspection. The pig eyeballs were immersed in 1% povidone-iodine solution for 2 minutes, rinsed with sterile PBS, immersed in PBS containing 0.1% gentamicin for 15 minutes, and keratectomy was performed after disinfection. In the complete eyeball, the separated cornea was resected, and 2-4mm of intact annular scl...
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Abstract
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