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Preparation method for wharton jelly tissue engineering scaffold of human umbilical cord

A technology of Wharton's jelly tissue and Wharton's jelly, applied in the field of biomedicine, can solve problems such as poor mechanical properties, unsuitable control of degradation rate, and uncertain elimination of antigenicity

Inactive Publication Date: 2019-04-26
康泽生医学生物科技(武汉)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Aiming at the deficiencies of the prior art, the present invention provides a preparation method of human umbilical cord Wharton's jelly tissue engineering scaffold, which solves the problems of poor mechanical properties, uncertain elimination of antigenicity and unsuitable control of degradation rate in existing scaffolds

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Specifically include the following steps:

[0025] S1. Separation of Wharton's jelly; remove blood vessels from the fresh umbilical cord, and then cut into pieces, Wharton's jelly tissue is derived from neonatal umbilical cord tissue;

[0026] S2. Mechanical pulverization: add the Wharton's jelly shredded in S1 to normal saline, and then shake to form a slurry;

[0027] S3. Repeated freezing and thawing: divide the S2 homogenate into cryopreservation tubes and freeze in liquid nitrogen for 24 hours, rewarm at 37 degrees Celsius for 1 hour, repeat 3 times;

[0028] S4. Differential speed centrifugation: centrifuge the frozen-thawed homogenate of S3 at 1800 g for 20 minutes, retain the supernatant, add 1% Tritin X-100, 0.25% trypsin, Tris-HCl buffer, and stir at 4 degrees Celsius for 24 hours. Centrifuge at 2500g for 30 minutes, collect the supernatant, add DNase and RNase, digest at 37°C for 16 hours, centrifuge at 3800g at 4°C for 20 minutes, adjust the pH to 7.0, take...

Embodiment 2

[0032] The scaffold was soaked in the culture medium for 20 min and then placed in a 6-well plate. Human umbilical cord mesenchymal stem cells were inoculated onto the scaffold at a density of 1×107 cells / ml, and incubated at 37°C for 2 h. After the cells adhered to the scaffold, add complete Culture medium 2-3ml / well, culture in 37 degrees Celsius, 5% CO2 cell incubator for 2-3 days, observe cell morphology and adhesion under electron microscope, it can be seen that the cells are in good growth state and have matrix secretion under the electron microscope.

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Abstract

The invention discloses a preparation method for a wharton jelly tissue engineering scaffold of human umbilical cord. The preparation method specifically comprises the following steps: S1) separatingwharton jelly; removing blood vessels from fresh umbilical cord and then cutting into pieces; S2) mechanically smashing: adding wharton jelly cut in step S1 into normal saline, and then uniformly shaking into slurry. The invention relates to the technical field of biomedicine. The preparation method for the wharton jelly tissue engineering scaffold of human umbilical cord is capable of preparing the wharton jelly tissue engineering scaffold of human umbilical cord without cells by adopting cell-removing, freeze-drying and cross-linking processes; ingredients, such as glycosaminoglycan and II type collagen, in wharton jelly are remained in the prepared tissue engineering scaffold; cells inoculated onto the scaffold are adhered to scaffold duct walls and are under excellent growth state; substances are secreted; an in vivo test proves that no immunoreaction exists; the product has a bright application prospect, so that mechanical properties of scaffold are greatly promoted, antigenicityis effectively eliminated and a controllable degradation rate is achieved.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a preparation method of a human umbilical cord Wharton's jelly tissue engineering scaffold. Background technique [0002] Tissue-engineered skin uses a three-dimensional scaffold as a carrier and is obtained by planting cells on the scaffold. An ideal artificial skin scaffold should meet the requirements of both material and structure. In terms of materials: 1. Allow cells to adhere to the surface and promote cell proliferation , to retain the function of differentiated cells; 2. Degradability, materials and degradation products are non-cytotoxic and will not cause inflammation; 3. Good biocompatibility; 4. Wide range of sources, low price, no risk of disease transmission, etc. Features, in terms of structure: 1. It has high porosity to provide enough space for cell adhesion, regeneration of extracellular matrix and cell diffusion. The pore structure can allow cells to distri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/36A61L27/50A61L27/56
CPCA61L27/3604A61L27/3687A61L27/3691A61L27/3834A61L27/50A61L27/56A61L2430/40
Inventor 姜大奎
Owner 康泽生医学生物科技(武汉)有限公司
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