Preparation method and application of bioengineering decellularized eye conjunctiva

[0030] Conjunctival tissue obtained by low-temperature fixer and trypan blue staining. By using a low-temperature fixer, the conjunctival tissue can be flattened and fixed on a low-temperature ice plate, and then the conjunctiva and subconjunctival tissue can be clearly distinguished by conventional trypan blue staining, and finally the epithelial layer is smooth and the tissue is flat. Conjunctival material.

[0031] 1. Protect the conjunctival tissue with protective solution throughout the whole process. The formula of the protective solution is: RPMI-1640 medium, add chondroitin sulfate 25g / L, low molecular dextran 10g / L, tobramycin 2.5mg / L, adjust pH value to 7.2, the osmotic pressure is 350mOsm;

[0032] 2. Seal the fresh conjunctiva after the subconjunctival tissue has been removed in a plastic bag filled with protective solution;

[0033] 3. Under the condition of 300MPa high static pressure, treat twice, each time is 2 minutes;

[0034] 4. After taking out the conju...

[0038] Conjunctival tissue obtained by low-temperature fixer and trypan blue staining. By using a low-temperature fixer, the conjunctival tissue can be flattened and fixed on a low-temperature ice plate, and then the conjunctiva and subconjunctival tissue can be clearly distinguished by conventional trypan blue staining, and finally the epithelial layer is smooth and the tissue is flat. Conjunctival material.

[0039] 1. Use the protective solution to protect the conjunctival tissue throughout the whole process. The formula of the protective solution is:

[0040] RPMI-1640 medium, add chondroitin sulfate 40g / L, low molecular weight dextran 20g / L, tobramycin 2.5mg / L, adjust the pH value to 7.2, and the osmotic pressure is 350mOsm;

[0041] 2. Seal the fresh conjunctiva after the subconjunctival tissue has been removed in a plastic bag filled with protective solution;

[0042] 3. Under the condition of 250MPa high static pressure, treat 5 times, each time is 1 minute;

[0043...

[0047] Conjunctival tissue obtained by low-temperature fixer and trypan blue staining. By using a low-temperature fixer, the conjunctival tissue can be flattened and fixed on a low-temperature ice plate, and then the conjunctiva and subconjunctival tissue can be clearly distinguished by conventional trypan blue staining, and finally the epithelial layer is smooth and the tissue is flat. Conjunctival material.

[0048] 1. Protect the conjunctival tissue with protective solution throughout the whole process. The formula of the protective solution is: RPMI-1640 medium, add chondroitin sulfate 50g / L, low molecular weight dextran 5g / L, tobramycin 2.5mg / L, adjust pH value to 7.2, the osmotic pressure is 380mOsm;

[0049] 2. Seal the conjunctiva after the subconjunctival tissue has been removed in a plastic bag filled with protective solution;

[0050] 3. Under the condition of 200MPa high static pressure, treat 3 times, each time is 2 minutes;

[0051] 4. After taking out the con...