A method for decellularization of human corneal stroma

Abstract

The invention aims to solve the problem of donor shortage of corneal stroma and overcome defects of a traditional decellularization method, provides a method for decellularization of human corneal stroma, and belongs to the technical field of cornea in tissue engineering. The method comprises the following steps: collecting corneal stroma containing tissue cells to be put in a cleaning solution, and washing in the cleaning solution for 2-3 times and 2-5 minutes each time; sealing the corneal stroma in nitrogen, and storing for 5-7 days; taking out the corneal stroma from nitrogen, washing by the cleaning solution for 2-3 times and 2-5 minutes each time, shaking at constant temperature in the cleaning solution, changing the cleaning solution every 1-2 hours to obtain decellularized corneal stroma. The method can be used for completely decellularization, collagen of corneal stroma cannot be hurt, preparations applied in the whole process are non-toxic, the biological characteristics of the corneal stroma can be furthest protected, and the stroma cells can be effectively removed.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering cornea, and in particular relates to a decellularization method of human corneal stroma. Background technique [0002] About 3 to 5 million Chinese citizens are blinded by monocular and binocular corneal diseases, ranking second only to cataracts. The main diseases that lead to corneal blindness include various infectious corneal diseases, corneal degeneration, severe mechanical trauma, ocular surface chemical injury, severe dry eye, complex pterygium, ocular pemphigoid infection, etc., covering almost all corneal diseases kind. The only way for most of these patients to obtain useful vision is surgical treatment, and the main method of surgery is full-thickness or lamellar corneal transplantation. Due to the limitation of traditional concepts and eye bank conditions in my country, the number of corneal donors is very limited, the treatment cost is high, and some corneal donors cannot...

AI Technical Summary

Problems solved by technology

Although there are many sources of corneal stroma material and decellularization methods, there are many problems: (1) corneal stroma material comes from human or animal eyes, and human corneal tissue compatibility is the best, but full-thickness corneal The sources are scarce, and the corneal stroma derived from animal eyes is difficult to avoid immune reactions and other problems; (2) In order to minimize the risk of immune reactions, the corneal stroma used for corneal transplantation needs to be decellularized in advance of the stromal cells on the corneal stroma

The development of the decellularization method from the initial freeze-thaw method to the more commonly used surfactant elution method has some unavoidable problems: for example, repeated freezing and thawing may cause collagen breakage and damage the tension of the corneal stroma; while surfactant All have relatively high toxicity, and it is difficult to apply clinically after treatment, and most of them stay in the laboratory stage at present.

[0005] There have been decades of research on corneal stroma decellularization methods. As mentioned earlier, many previous decellularization methods were difficult to promote clinically due to their limitations.