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Preparation of human umbilical cord mesenchymal stem cells wound surface smearing agent and storage application method

A technology of mesenchymal stem cells and human umbilical cord, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc.

Inactive Publication Date: 2011-04-06
戴育成
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  • Abstract
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Problems solved by technology

Although artificial composite skin has been used to treat skin injuries in recent years, the thin-layer full-thickness graft is an imperfect full-thickness skin substitute, which has the following problems: 1. The artificial composite skin is thick and has poor flexibility, Wounds that cannot fully adhere to uneven skin
However, the biggest problem in the application of umbilical cord blood is that there are many umbilical cord blood samples whose mononuclear cells cannot form mesenchymal stem cells, so its popularization and application are greatly limited.
In recent years, it has been found that most umbilical cord Wharton's jelly can form mesenchymal stem cells, but the separation and purification methods or separation steps reported so far are many and time-consuming, or the degree of purification of mesenchymal stem cells is not high

Method used

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Embodiment Construction

[0053] The present invention is described in further detail below in conjunction with embodiment.

[0054] 1. A method for separating, purifying and culturing human umbilical cord mesenchymal stem cells:

[0055] a. With the consent of the family members, soak the umbilical cord of a normal full-term fetus in 5% chlorhexidine for 10 minutes, wash the blood vessels in the umbilical cord with chlorhexidine at the same time, remove the blood in the blood vessels, and cut the umbilical cord into 3 parts with surgical scissors. - 4 cm long section, soaked in a 50ml centrifuge tube containing 5% chlorhexidine for 5 minutes, then soaked and rinsed with normal saline for 3 times, 3 minutes each time;

[0056] b. Cut each section of umbilical cord along the longitudinal axis, peel off the amniotic epithelial layer on the surface, expose the umbilical artery and umbilical vein, peel off and extract the umbilical artery and umbilical vein, and then perform the remaining Wharton's jelly t...

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Abstract

The invention discloses a method for separating, purifying and culturing human umbilical mesenchymal stem cells and a method for preparing, storing and quickly preparing and applying a wound surface painting reagent of the human umbilical mesenchymal stem cells. The method comprises: firstly, separating, purifying and culturing the human umbilical mesenchymal stem cells; secondly, preparing a methyl cellulose composite culture medium; thirdly, preparing the wound surface painting reagent of the human umbilical mesenchymal stem cells; and fourthly, detecting the biological properties of the human umbilical mesenchymal stem cells. The method takes methyl cellulose as a substrate, mixes the human umbilical mesenchymal stem cells into the methyl cellulose, and generates the wound surface painting reagent of the human umbilical mesenchymal stem cells, and the wound surface painting reagent can be uniformly coated on the wound surface of the skin to form a coating. The invention performs preclinical researches such as animal toxicity test, pyrogen test and hypersensitive test, constructs a cryopreservation resuscitation method for human umbilical mesenchymal stem cells and a stem cell bank, and prepares for large-scale production and preparation of the wound surface painting reagent of the human umbilical mesenchymal stem cells and application of the wound surface painting reagent of the human umbilical mesenchymal stem cells.

Description

technical field [0001] The invention relates to burn medicine, in particular to a preparation, storage and application method of a human umbilical cord mesenchymal stem cell wound varnish. Background technique [0002] Traditional methods of treating skin injuries such as skin wounds and burns are not ideal. Although artificial composite skin has been used to treat skin injuries in recent years, the thin-layer full-thickness graft is an imperfect full-thickness skin substitute, which has the following problems: 1. The artificial composite skin is thick and has poor flexibility, It cannot be completely attached to the uneven wound surface of the skin. After covering the wound with it, the secretion of the wound can make the composite skin bulge; 2. The seed cells of the artificial composite skin currently used are generally autologous or allogeneic epidermal cells, and the source of autologous epidermal cells is sometimes insufficient; while allogeneic epidermal cells invol...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/08A61K35/44A61K9/70A61P17/02A61K35/51C12N5/0775
Inventor 戴育成胡葵葵董凤平
Owner 戴育成
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