Tissue engineered esophagus

A tissue engineering and esophagus technology, applied in medical science, prosthesis, etc., can solve the problems of unsatisfactory biocompatibility and cell affinity, and achieve good cell compatibility and good degradation rate.

Inactive Publication Date: 2013-06-19
XIAN REJE BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the past, polyglycolic acid and other synthetic polymer degradable materials were often used, but their biocompatibility and cell affinity were not ideal

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Isolation and culture of epidermal keratinocytes

[0019] The patient's epidermis was trimmed into strips with a width of 1-2 stomachs, rinsed twice in PBS without calcium and magnesium, and immersed in 0.5% neutral protease overnight with the epidermis facing upward. After the epidermis was torn off from the dermis, it was cut into pieces and digested with 0.05% trypsin-0.53mM EDTA at 37°C for 20 minutes, and 10ml of trypsin inhibitor was used to terminate the digestion. Filter through a 150-mesh stainless steel filter, centrifuge (1200 rpm, 10 minutes) and count to make a single-cell suspension, press 10 5 / m 2 Density seeding culture, serum-free epidermal cell medium, plus bovine pituitary extract and recombinant epidermal growth factor, at 37°C, 5% CO 2 Cultured in an incubator with saturated humidity, the culture medium was changed every 3 days. When the cells reached 60%-70% confluence, they were digested with 0.05% trypsin at 37°C for 10 minutes, and subcultur...

Embodiment 2

[0021] Dermal Fibroblast Isolation and Culture Isolation and Digestion

[0022] The leather fragments in Example 1 were digested with 2% collagenase at 37°C for 2 hours, filtered through a 150-mesh stainless steel filter, centrifuged at 1200 rpm for 5 minutes, the supernatant was discarded, and calcium and magnesium-free PBS was added to wash 3 times. Add 10ml of culture medium containing 10% fetal bovine serum, mix well, and count by trypan blue staining.

[0023] After counting the primary culture, make a single cell suspension, press 10 5 / cm 2 Density inoculation culture, DMEM culture solution containing 10% fetal bovine serum, cultivated in an incubator at 37°C and 10% relative humidity, and the culture solution was changed every 3 days. When the cells are confluent to 60%-75% density, wash with 1% calcium and magnesium-free PBS, digest with 0.25% trypsin at 37°C for 5 minutes, add DMEM medium containing 10% fetal bovine serum, mix well, count, press Subculture at a r...

Embodiment 3

[0025] Constructing tissue engineered esophagus

[0026] Vascular endothelial cells and dermal fibroblasts cultured in vitro were collected, seeded on tissue-engineered skin scaffold materials, and vascular endothelial cells, fibroblasts-acellular dermal complexes were constructed. Seeding density is 10 5 -10 8 . The culture medium was changed every 3 days, and the culture time was 7-10 days.

[0027] Form a double-layer structure: collect epidermal cells and inoculate them on the surface of vascular endothelial cells, fibroblasts-degradable scaffold material composites. After one week, an air-liquid interface formed on the surface of the complex, promoting further differentiation of vascular endothelial cells.

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PUM

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Abstract

The invention discloses a tissue engineered esophagus and a preparation method thereof. The tissue engineered esophagus contains a dual-layer structure of an epithelial layer and a dermis layer constructed by a scaffold material and seed cells, wherein the scaffold material is acellular matrix, and the seed cells include fibroblasts, vascular endothelial cells, and keratinocytes. The vascular endothelial cells in the dermis layer directly take part in the creation of capillary network, and two revascularization methods are used for quickly communicating with the blood supply channel of receptor, thus the epithele of the tissue engineered esophagus is provided with blood.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering, and in particular relates to a tissue-engineered esophagus and a preparation method thereof. Background technique [0002] Tissue engineering applies the principles and methods of life science and engineering to research and develop biological substitutes for repairing, enhancing or improving the functions and shapes of various human tissues or organs after injury. The basic preparation method of the biological substitute is to combine cells with degradable biomaterials after in vitro expansion to form a cell-degradable biomaterial complex to replace damaged tissues and organs. It has the characteristics of forming living tissues with vitality, repairing large defects with a small number of cells, and can be shaped arbitrarily according to the defects of tissues and organs. As far as the tissue-engineered heart esophagus is concerned, there are still two technical difficulties in the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/40A61L27/38A61L27/36
Inventor 步怀宇
Owner XIAN REJE BIOLOGICAL TECH
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