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Nano-polypeptide hydrogel of composite stem cell spheres as well as preparation method and application of nano-polypeptide hydrogel

A technology of stem cell spheres and hydrogels, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve problems such as failure to meet clinical needs, secondary damage to the donor site, and limited effectiveness, and help the epidermis Reconstruction and vascular regeneration, easy survival, pain relief effect

Pending Publication Date: 2022-06-24
QINGDAO WOMEN & CHILDREN HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Various acute and chronic wounds or ulcers are formed on the skin due to various reasons such as severe trauma, extensive burns, and diabetes, and the damaged skin cannot be healed quickly and perfectly
[0003] Large skin defects are usually repaired by skin grafting or flap transplantation, but autologous skin grafting causes secondary damage to the donor site
Therefore, people have to focus on artificial skin substitutes, but the clinically available artificial skin has problems such as immunogenicity, its effectiveness is limited, and it is far from meeting clinical needs

Method used

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  • Nano-polypeptide hydrogel of composite stem cell spheres as well as preparation method and application of nano-polypeptide hydrogel
  • Nano-polypeptide hydrogel of composite stem cell spheres as well as preparation method and application of nano-polypeptide hydrogel
  • Nano-polypeptide hydrogel of composite stem cell spheres as well as preparation method and application of nano-polypeptide hydrogel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] This example is the isolation and culture of human umbilical cord mesenchymal stem cells, and the formation of stem cell spheres:

[0045] Fresh human umbilical cord tissue was obtained in the clinic, and the umbilical cord tissue-derived mesenchymal stem cells were isolated by collagenase digestion. The medium was routinely cultured. After the cells grew to 80% confluence, trypsin digestion was used to digest the adherent cells; selection (FACS) for identification.

[0046] The resulting human umbilical cord mesenchymal stem cells (UC-MSCs) were grown in high glucose Dulbecco's modified Eagle's medium supplemented with 20% fetal bovine serum, and incubated in 5% CO. 2 Culture in an incubator at 37.0°C, passage every 5-7 days; obtain UC-MSCs between passages 6-10 for experiments.

[0047] Formation of stem cell spheres:

[0048] Formation of UC-MSCs spheroids (UC-MSCsp) using the hanging drop method: UC-MSCs were isolated and separated at 1 × 10 6 UC-MSCsp were harve...

Embodiment 2

[0056] This embodiment is a preparation method of self-assembled polypeptide hydrogel, and the preparation method includes the following steps:

[0057] Polypeptides RADA16-I, RGD and KLT were dissolved in sterile ultrapure water by ultrasonic respectively to obtain an aqueous solution of polypeptide RADA16-I with a concentration of 10 g / L, an aqueous solution of polypeptide RGD with a concentration of 10 g / L and a polypeptide with a concentration of 10 g / L. KLT aqueous solution;

[0058] Mix the polypeptide RADA16-I aqueous solution, the polypeptide RGD aqueous solution and the polypeptide KLT aqueous solution according to the volume ratio of 2:1:1 to obtain the polypeptide mixture. The basement membrane of the 24-well plate is submerged in the wells of the 24-well plate; 100ul of the polypeptide mixture is slowly added to the insert in each cross-well plate against the wall, so that the PBS in the 24-well plate induces the self-assembly of the polypeptide mixture in the inse...

Embodiment 3

[0066] This embodiment is a preparation method of nano-polypeptide hydrogel of composite stem cell sphere, and the preparation method includes:

[0067] According to the seeding density of 40 stem cell spheres per 30 μl of self-assembled polypeptide hydrogels, the stem cell spheres in Example 1 were inoculated into the self-assembled hydrogels in Example 2; that is, the nanopolypeptides of the composite stem cell spheres were obtained. Hydrogels.

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Abstract

The invention discloses nano polypeptide hydrogel of composite stem cell spheres as well as a preparation method and application of the nano polypeptide hydrogel. The preparation method comprises the following steps: (a) preparing the stem cell spheres; (b) preparing the self-assembled polypeptide hydrogel; and (c) compounding of the stem cell spheres and the self-assembled polypeptide hydrogel: inoculating the stem cell spheres into the self-assembled hydrogel to obtain the nano polypeptide hydrogel compounded with the stem cell spheres. According to the invention, the human umbilical cord mesenchymal stem cell spheres and the nano-polypeptide hydrogel are combined and can be directly smeared on a skin wound, so that the survival of transplanted cells is greatly improved, the pain in the transplanting process is relieved, and the operative wound is avoided; the transplanted stem cells gradually disappear after wound healing, and the method is more convenient, safer and more efficient.

Description

technical field [0001] The invention relates to the technical field of biological materials, in particular to a nano-polypeptide hydrogel of composite stem cell spheres and a preparation method and application thereof. Background technique [0002] Various acute and chronic wounds or ulcers are formed on the skin due to various reasons such as severe trauma, extensive burns and diabetes, which cannot complete the rapid and perfect healing of the damaged skin. [0003] Large-scale skin defects are usually repaired by skin grafting or skin flap transplantation, but autologous skin grafting results in secondary damage to the donor site. Therefore, people have to focus on artificial skin substitutes, but the clinically available artificial skin has problems such as immunogenicity, its effectiveness is limited, and it is far from meeting clinical needs. At present, the application of tissue-engineered skin and stem cell-related products has attracted more and more attention. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K9/06A61K35/28A61K47/42A61P17/02
CPCC12N5/0668A61K9/06A61K35/28A61K47/42A61K9/0014A61P17/02C12N2533/50C12N2535/00
Inventor 孙念峰刘军帅田爱玲伊鑫
Owner QINGDAO WOMEN & CHILDREN HOSPITAL
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