Preparation and application of cell-eliminating pig corium substrate without cytotoxicity

Abstract

The preparation process of cell-eliminating pig corium substrate includes sustained vibration of the corium substrate in 0.5 mol / L NaOH solution compounded with thrice distilled water, rinsing with thrice distilled water and balanced phosphate salt solution separately, freeze drying, packing and sterilizing. The cell-eliminating pig corium substrate is used as the in vitro corium culturing rack in composite skin preparing tissue engineering process, and the preparation process includes coating the corium substrate with acetic acid dissolved type-I ox collagen, regulating pH to 7.0-7.4, soaking subsequently in balanced phosphate salt solution and no serum culture medium, inoculating passage epidermal cell, and no serum culture inside a CO2 hatcher. The tissue engineering composite skin is used as human skin substitute.

Description

Technical field [0001] The invention belongs to the technical field of medical skin tissue engineering, in particular to a method for preparing a non-cytotoxic pig acellular dermal matrix; relates to a method for constructing a tissue engineering composite skin with a non-cytotoxic pig acellular dermal matrix as a scaffold; relates to a non-cytotoxic pig Cellular dermal matrix is ​​used as a skin substitute in the construction of tissue engineering composite skin and tissue engineering composite skin preparation. Background technique [0002] At present, the preparation methods of porcine acellular dermal matrix as a substitute for human skin mainly include enzyme digestion method, hypertonic salt-detergent treatment method and balanced salt solution method. Among them, the enzyme digestion method has been reported most at home and abroad. It uses exogenous proteases to remove the epidermal structure at 4°C or 37°C, supplemented by other methods, such as tissue fluid soaking and ...

AI Technical Summary

Problems solved by technology

In the above two methods, some chemical reagents used in the preparation, such as Tritonx-100, SDS, etc., have a strong cell lysis effect, which is difficult to completely remove, resulting in a low success rate of cell culture using the dermal matrix as a scaffold, and poor reproducibility; balance The saline solution method is to place sterile skin pieces in 37°C phosphate-buffered saline solution to activate the endogenous proteases of the skin, thereby separating the epidermis and dermis, and then remove all the memory of the dermis by repeated freeze-thawing and gamma-ray irradiation. Living cells

But this method, first, the production process is relatively complicated, and the processing time is long, about 2 weeks; second, it is easy to be polluted during the preparation process; third, the antigenicity is not completely removed