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
View PDF7 Cites 7 Cited by
  • 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, su

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 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

Example Embodiment

[0035] Example 1 ： Method for preparing double-layer skin containing microvascular lumen

[0036] Step 1. Extraction of fibroblasts and epidermal cells:

[0037] Dip the skin tissue in 75% ethanol solution, then transfer it to PBS solution to soak until there is no blood stain; add the tissue block to Dispase digestion solution, place it at 4℃ for 16 h; then add collagenase and incubate at 37℃ for 1 After stopping the digestion, centrifuge at 1000 rpm / min for 8 minutes, add fibroblast culture medium for culture, put the peeled epidermis into 8 ml of 0.25% pancreatin / EDTA digestion solution preheated at 37℃, and then incubate at 37℃ Digest in the box for 8 minutes, stop after the digestion is complete, filter the obtained epidermis, centrifuge at 800 r / min for 5 min; add PBS buffer after centrifugation, centrifuge at 800 r / min for 5 min; pour out the supernatant and add the epidermal cell culture solution , Carry out inoculation culture;

[0038] Step 2: Primary extraction of umbi...

Example Embodiment

[0054] Example 2 ： Method for preparing double-layer skin containing microvascular lumen

[0055] Step 1. Extraction of fibroblasts and epidermal cells:

[0056] Dip the skin tissue in 75% ethanol solution to remove blood stains, then transfer to PBS solution to soak until no blood stains; add the tissue block to Dispase digestion solution, place it at 4℃ for 17 h; then add collagenase and incubate at 37℃ Incubate in the box for about 2 hours. After terminating the digestion, centrifuge at 1000 rpm / min for 10 minutes and add fibroblast culture solution for culture; put the peeled epidermis into the 0.25% pancreatin / EDTA digestion solution preheated at 37°C and transfer to 37°C Digest in an incubator for 10 minutes, filter the obtained epidermis after termination, and centrifuge at 800 rpm / min for 5 minutes;

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

Example Embodiment

[0074] Example 3 ： Method for preparing double-layer skin containing microvascular lumen

[0075] Step 1. Extraction of fibroblasts and epidermal cells:

[0076] Dip the skin tissue in 75% ethanol solution to remove blood stains, then transfer to PBS solution for soaking; add the tissue block to Dispase digestion solution, place it at 4℃ for 16 h; then add collagenase to 37℃ incubator for incubation 2 About h, after terminating the digestion, centrifuge at 1000 rpm / min for 10 minutes, add fibroblast culture medium for culture; put the peeled epidermis into the 0.25% pancreatin / EDTA digestion solution preheated at 37℃, and transfer it to the 37℃ incubator Digest for 10 min, filter the obtained epidermis after termination, and centrifuge at 800 rpm / min for 5 min;

[0077] Pour out the supernatant after centrifugation, add 10 mL of PBS buffer, and centrifuge at 800 rpm / min for 5 min; after centrifugation, pour out the supernatant, add epidermal cell culture solution, and inoculate cu...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A61L27/60A61L27/38C12N5/071C12N5/0775
Inventor 孙蓓卢永波
Owner GUANGDONG BOXI BIO TECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap