Culture of tissue engineering skin and detection method of proliferation activity

A tissue-engineered skin, vitality detection technology, applied in tissue culture, biochemical equipment and methods, biological testing, etc., can solve the problems of skin quality and safety, affecting skin tissue growth and development, interference and other issues

Inactive Publication Date: 2009-03-25
陆洪光
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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 tissu

Method used

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Embodiment Construction

[0013] Embodiments of the present invention: the basic scheme of the present invention is as follows: firstly make a three-dimensional matrix material containing basement membrane components as a dermis substitute, i.e. human de-epidermized humandermis (HDED for short), and cultivate keratinocytes; Then a certain amount of keratinocytes were inoculated on the surface of HDED coated with tissue glue, and the organ culture was carried out by means of air-liquid surface culture. The culture was continued for about 12-15 days, and samples were taken with a micro-biopsy device to detect the skin proliferation activity. The rest of the skin tissue continued to be cultured, and the culture was terminated after continuing to culture for one week, the skin was taken out, and the skin proliferation activity was tested again. Place the freshly removed culture specimens in BrdU culture medium (protected from light) with a final concentration of 100 μg / ml, 37? After incubating in the incub...

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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...

Claims

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Application Information

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IPC IPC(8): C12N5/08A61L27/60G01N33/53G01N15/10C12N5/071
Inventor 陆洪光
Owner 陆洪光
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