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Tissue engineering skin and application thereof

A tissue-engineered skin and umbilical cord technology, applied to tissue-engineered skin and its application fields, can solve the problems of limited effect of repairing skin wounds, and achieve the effects of repairing skin wounds, fewer operation steps, and improving the number and proliferation ability.

Active Publication Date: 2015-06-03
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Dermal fibroblasts as seed cells can promote the proliferation and differentiation of epidermal stem cells, which is beneficial to wound healing, but the effect of repairing skin wounds is limited

Method used

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  • Tissue engineering skin and application thereof
  • Tissue engineering skin and application thereof
  • Tissue engineering skin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The construction of embodiment 1 tissue engineered skin

[0047] Coat 2×2 cm ADM with 7 mg / mL gelatin solution for 30 minutes, discard the remaining gelatin solution, and wash once with PBS. Add stem cell-specific medium (Lonza) to submerge ADM, discard the medium after 2 hours, and wipe the surface of ADM with sterile filter paper to absorb the surface liquid and keep the surface moderately moist.

[0048] Use a sterile stainless steel ring of the same size as the ADM to press on the periphery of the ADM, 5×10 6 Add human umbilical cord MSC (P1-P5 generations) suspension evenly on the surface of ADM until the surface of ADM is filled with liquid at a cell density of / mL, plant after 1 hour, and plant continuously for 3 times. As for the culture medium, lightly insert a fine needle into ADM to make several wells, and culture in a standard incubator (37°C). On the third day, a small piece of tissue was cut out for frozen section, and HE staining was performed for ident...

Embodiment 2

[0050] Example 2 Verification of Cell Planting Efficiency

[0051] The gelatin solution that is 3mg / mL, 5mg / mL, 7mg / mL, 9mg / mL is used to coat the ADM of 1cm * 1cm size with the concentration respectively, uses the cell planting method that embodiment 1 provides respectively in gelatin-coated ADM and uncoated ADM. Plant the cells on the surface of the ADM, collect the medium after 2 hours, and digest the cells attached to the wall of the bottle with 0.25% trypsin-0.53mM EDTA solution, calculate the amount of cells that are not attached to the ADM after planting, using the following formula Calculate cell seeding efficiency:

[0052] Cell planting efficiency = (total amount of planted cells - amount of cells not attached to ADM) / total amount of planted cells × 100%.

[0053] The results of cell planting efficiency in each group are shown in Table 1:

[0054] Table 1 Comparison of cell planting efficiency in each group (%, )

[0055] group

Cell Planting Effic...

Embodiment 3

[0058] Embodiment 3 tissue engineered skin transplantation

[0059] (1) Making a Rat Burn Model

[0060] 15 female SD rats, with a body weight of about 200 grams, were anesthetized by intraperitoneal injection of 3% pentobarbital sodium 30mg / kg the day before the burn, and scissors cut off the hair of the scalded part (located on the back), and used 10% vulcanization Sodium solution for hair removal. The next day, after the rats were anesthetized, they were injected intraperitoneally with tramadol injection (10 mg / kg) to relieve pain, and a red copper rod with a diameter of 2 cm, a length of 10 cm, and a temperature of 100 ° C (heated in boiling water at 100 ° C for more than 10 minutes) was used in the large Three burns were made on the back of the rats (burning time was 12 seconds), and the degree of burns was confirmed as third-degree burns by histopathological examination. Immediately after the burn, 2 mL of lactated Ringer's solution was injected intraperitoneally to ai...

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Abstract

The invention relates to the field of tissue engineering, in particular to tissue engineering skin and an application thereof. A construction method of the tissue engineering skin comprises steps as follows: umbilical cord mesenchymal stem cells are adopted as seed cells, an ADM (acellular dermal matrix) serves as a support, and the tissue engineering skin is constructed and obtained. The tissue engineering skin can restore skin wounds more effectively, and the construction method of the tissue engineering skin is simple and convenient and comprises few operation steps.

Description

technical field [0001] The invention relates to the field of tissue engineering, in particular to a tissue-engineered skin and its application. Background technique [0002] The skin is the first barrier for the human body to resist external damage. The skin is the largest and most complex organ in the human body, and it is also the most vulnerable organ in the process of burn trauma. Large-scale skin defects can cause loss of body fluids, water and electrolyte disorders, and hypoproteinemia. disease, severe infection, etc., and skin transplantation is the key to solve this problem, but because the sources and applications of autologous and allogeneic skin are limited in some cases, people have been looking for ideal skin substitutes. Skin tissue engineering is one of the promising ways to solve this problem. Skin tissue engineering is an emerging marginal subject, and it is currently the closest to successful histological products. It uses the principles of biology and en...

Claims

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Application Information

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IPC IPC(8): A61L27/60A61L27/36
Inventor 陈海佳王一飞葛啸虎戴国胜王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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