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Tissue-engineered epidermis quality detection method

A tissue engineering and detection method technology, applied in biochemical equipment and methods, biological testing, measuring devices, etc., can solve the problems of lack of quality and safety detection methods for tissue engineering epidermis

Inactive Publication Date: 2015-12-09
赫柏慧康生物科技无锡有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the current lack of systematic tissue engineering epidermis quality and safety testing methods, the present invention provides a tissue engineering epidermis testing method

Method used

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  • Tissue-engineered epidermis quality detection method
  • Tissue-engineered epidermis quality detection method
  • Tissue-engineered epidermis quality detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Take the foreskin skin of (human) healthy male after circumcision operation, and use an area of ​​2cm 2 Keratinocytes and melanocytes were extracted and cultured from the left and right skin samples, and this batch of tissue engineered skin into skins in the watch glass. A piece of skin was randomly selected for detection of melanocytes and uniformity of the skin, and all skins were tested for biological safety. The implementation method and results are as follows:

[0038] (1) Preparation of DOPA dye solution

[0039] Use phosphate buffer solution with a pH of 7.2-7.6 to prepare DOPA powder to a final concentration of 0.1%, and stir on a magnetic stirrer at room temperature for 30 minutes to dissolve.

[0040] (2) DOPA staining

[0041] A piece was randomly selected from the cultured tissue engineered epidermis, fixed with 4% formaldehyde for 10 minutes, rinsed with phosphate buffer solution with a pH of 7.2-7.6, dried overnight, and then stained with 0.1% DOAP in...

Embodiment 2

[0058] Take the foreskin skin of (human) healthy male after circumcision operation, and use an area of ​​3cm 2 Keratinocytes and melanocytes were extracted and cultured from the left and right skin samples. This batch of tissue-engineered skin into skins in the watch glass. A skin slice was randomly selected for detection of melanocytes and keratinocytes, and all skin slices were subjected to biosafety testing. The detection method is the same as in Example 1. The result is as follows:

[0059] (1) The results of DOPA staining and hematoxylin staining are as follows image 3 , the picture shows that although there are melanocytes (pointed by black arrows), the distribution is uneven, and the distribution of keratinocytes is severely uneven (pointed by triangles).

[0060] (2) Sterility testing

[0061] The sterility test results after 24 h of cultivation are shown in Table 3. All skin grafts were tested negative for sterility.

[0062] Table 3, Sterility test results ...

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Abstract

The invention provides a tissue-engineered epidermis detection method. The tissue-engineered epidermis detection method includes melanocyte and keratinocyte detection, sterile detection, mycoplasma detection and endotoxin detection. By means of staining of cell nucleuses of melanocytes and keratinocytes, distribution of the melanocytes and the keratinocytes can be seen, so that melanocyte uniformity, skin graft uniformity and a proportion of the melanocytes and the keratinocytes on a basal layer can be seen; biosafety of tissue-engineered epidermis is detected by sterile detection, mycoplasma detection, endotoxin detection and the like. Thus, the tissue-engineered epidermis detection method can be used for detecting whether content of the melanocytes on the basal layer, skin graft uniformity and biosafety of the tissue-engineered epidermis reach the standards or not objectively, comprehensively and accurately.

Description

technical field [0001] The invention relates to a method for detecting the quality of tissue engineering epidermis, in particular to a method for detecting the uniformity and safety of melanin and keratinocytes. Background technique [0002] Wound is the loss of skin integrity due to internal or external factors. Its repair has always been the main problem in clinical practice. According to time, clinical wounds can be divided into acute wounds and chronic wounds. Acute wounds include burns, trauma and other major diseases, or skin defects formed after surgical resection of scars, skin pigmented diseases, benign and malignant tumors, etc.; chronic wounds include traumatic ulcers, diabetic ulcers, vascular ulcers, pressure ulcers, radiation Ulcers, tumorous ulcers, cicatricial ulcers, etc. [0003] At present, the most effective method for repairing skin defects is autologous skin transplantation, but autologous skin sources for large-area burns often cannot meet the needs o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/22C12Q1/68G01N21/78G01N21/17G01N33/50
Inventor 刘洪李琳杨熙
Owner 赫柏慧康生物科技无锡有限公司
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