Culture of tissue engineering skin and detection method of proliferation activity

Abstract

The invention discloses a detection method of tissue engineering skin culture and proliferation activity, which comprises the following process: first three-dimensional host materials containing basilar membrane are prepared as the substitute of dermis, namely dermis tissue of human with epidermis being removed, and keratinocyte is cultivated; the keratinocyte is inoculated to the surface of the dermis tissue of human with epidermis being removed and coated with mixed tissue glue, sampling is carried out for the detection of the skin proliferation activity after the culture of air-liquid surface culture mode, the culture is stopped after the culture is continued for a week, and then the skin is picked out for skin proliferation activity detection. A fresh culture specimen picked is put into BrdU culture liquid and a culture box of 37 DEG C for breeding, immnohistochemical staining is carried out after 24 hours, proliferation cells are observed under a microscope, the quantity of proliferation cells is calculated by using a computer imaging system, and tissue engineering skin structure is observed by the method of histology simultaneously. The detection method of tissue engineering skin culture and proliferation activity can detect the impact of different culture conditions and relevant factors on the tissue engineering skin cell proliferation by quantization.

Description

technical field [0001] The invention relates to a method for detecting tissue engineering skin culture and proliferation activity. Background technique [0002] Tissue engineered skin can be used for skin transplantation after skin trauma such as injury, burn, etc., cosmetic skin resurfacing and treatment of some serious skin diseases. It has broad application prospects and potential development space (Ehrenreich M, Ruszczak Z. Update on tissue- engineered biological dressings. Tissue Eng, 2006, 12: 2407-2424.). In the process of culturing tissue-engineered skin, maintaining the viability of seed cell proliferation is the key to skin development, and it is also an important reference for judging whether the skin can survive after transplantation. Therefore, in order to promote the proliferative activity of tissue engineered skin cells, people continue to explore appropriate culture conditions and add various biological components to the medium, such as bovine pituitary extr...

AI Technical Summary

Problems solved by technology

However, since BrdU is an exogenous cell marker, if BrdU is added to the whole cultured tissue engineered skin to detect its proliferation, it will interfere with and affect the growth and development of skin tissue, and the BrdU labeling of living tissue may cause tissue damage. The quality of engineered skin and the safety of its subsequent use in human skin transplantation will be affected, so it is necessary to find an ideal method that can objectively detect the proliferation of tissue-engineered skin without affecting skin growth