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Dual-layer skin with capillary lumina and preparation method of dual-layer skin

A technology of microvessels and lumens, applied in the field of double-layer skin and its preparation, can solve the problems of complexity and instability in the construction process, and achieve the effects of shortening the culture time, easy acquisition, and increasing the thickness

Active Publication Date: 2017-04-05
GUANGDONG BOXI BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The commonly used added factors are: vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF), etc., this method often requires repeated implementation or strict control of the release system, such as added The concentration of factors, the ratio of different added factors, and the action time of each factor, etc., all inevitably lead to the complexity and instability of the entire construction process, which limits its application to a certain extent.

Method used

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  • Dual-layer skin with capillary lumina and preparation method of dual-layer skin
  • Dual-layer skin with capillary lumina and preparation method of dual-layer skin
  • Dual-layer skin with capillary lumina and preparation method of dual-layer skin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 ： A preparation method of double-layer skin containing microvascular lumen

[0036] Step 1, the extraction of fibroblasts and epidermal cells:

[0037] Immerse the skin tissue in 75% ethanol solution, and then transfer it to PBS solution until there is no blood stain; add the tissue block to Dispase digestion solution, place it at 4°C for 16 h; then add collagenase and incubate in a 37°C incubator for 1 About h, after terminating digestion, centrifuge at 1000rpm / min for 8min, add fibroblast culture medium for culture, put the stripped epidermis into 8 ml 37℃ preheated 0.25% trypsin / EDTA digestion solution, and transfer to 37℃ for incubation Digest in the box for 8 minutes, stop after the digestion is completed, filter the obtained epidermis, and centrifuge at 800 r / min for 5 minutes; add PBS buffer after centrifugation, and centrifuge at 800 r / min for 5 minutes; pour off the supernatant and add epidermal cell culture medium , carry out inoculation culture...

Embodiment 2

[0054] Example 2 ： A preparation method of double-layer skin containing microvascular lumen

[0055] Step 1, the extraction of fibroblasts and epidermal cells:

[0056] Immerse the skin tissue in 75% ethanol solution to remove blood stains, then transfer to PBS solution for rinsing until there is no blood stain; add the tissue block to Dispase digestion solution, place it at 4°C for 17 h; add collagenase and incubate at 37°C Incubated in the box for about 2 hours, centrifuged at 1000rpm / min for 10min after the digestion was terminated, and added fibroblast culture medium for cultivation; the peeled epidermis was placed in 0.25% trypsin / EDTA digestion solution preheated at 37°C, and transferred to 37°C Digest in the incubator for 10 min, then filter the epidermis obtained after termination, and centrifuge at 800 rpm / min for 5 min;

[0057] Pour off the supernatant after centrifugation, add 10 mL PBS buffer solution, and centrifuge at 800 rpm / min for 5 min; pour off the sup...

Embodiment 3

[0074] Example 3 ： A preparation method of double-layer skin containing microvascular lumen

[0075] Step 1, the extraction of fibroblasts and epidermal cells:

[0076] The skin tissue was immersed in 75% ethanol solution to remove blood stains, and then transferred to PBS solution for soaking; the tissue block was added to Dispase digestion solution, and placed at 4°C for 16 hours; then collagenase was added and incubated in a 37°C incubator for 2 About h, after terminating digestion, centrifuge at 1000rpm / min for 10min, add fibroblast culture medium for culture; put the stripped epidermis into 0.25% trypsin / EDTA digestion solution preheated at 37°C, and transfer it to a 37°C incubator After digestion for 10 min, the epidermis obtained by filtration was terminated, and centrifuged at 800 rpm / min for 5 min;

[0077] Pour off the supernatant after centrifugation, add 10 mL PBS buffer solution, and centrifuge at 800 rpm / min for 5 min; pour off the supernatant after centrifu...

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Abstract

The invention provides a dual-layer skin with capillary lumina. The dual-layer skin is provided with two skin texture layers which are the epidermal layer and the corium layer from top to bottom, and the corium layer contains the capillary lumina. The epidermal layer is composed of multiple epidermic cell germinal layers. The corium layer containing the capillary lumina is formed by fiber cells, umbilical cord mesenchymal stem cells and umbilical vein endothelial cells, specifically, the fiber cells, the umbilical cord mesenchymal stem cells and the umbilical vein endothelial cells are mixed according to a certain proportion; the inoculation method is adopted repeatedly; and through the extracellular matrix secretion capacity and paracrine system of the cells, a skin corium histological structure is formed on the one hand, and on the other hand, multiplication and migration of endothelial cells and lumen formation and stabilization are prompted. By means of the texture construction method which simulates normal skin composition of human beings and achieves constitution of the corium layer and formation of the micro lumina in the corium through interaction of the cells, structure and function instability and operation complexity caused by manual addition of exogenous stents and angiogenic factors are avoided.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering in biomedical engineering, and mainly relates to a double-layer skin containing microvascular lumens and a preparation method thereof. Background technique [0002] The demand of normal skin cells for oxygen and nutrients and the removal of metabolic wastes are realized by blood diffusion and transportation near the cells, but the current double-layer skin based on fibroblast composite scaffold materials and epidermal cells cannot meet the requirements due to the lack of vascular structure. Long-term survival and functional requirements lead to dysfunction of fibroblasts, reduced migration and proliferation of keratinocytes, and ultimately limit the function of skin tissue; The acquisition of nutrients and waste excretion of living cells in the double skin mainly depends on the exudate of the wound base, so the material exchange speed of the cells is slow, which seriously affects the su...

Claims

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Application Information

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IPC IPC(8): A61L27/60A61L27/38C12N5/071C12N5/0775
Inventor 孙蓓卢永波
Owner GUANGDONG BOXI BIO TECH CO LTD
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