Supernatant gel stem cell culture medium for repairing bone injury and preparation method thereof

A damage repair and stem cell technology, applied in cell dissociation methods, cell culture active agents, bone/connective tissue cells, etc., can solve problems such as lack of directional differentiation efficiency, and achieve significant results in eliminating the risk of secondary bone fractures

Pending Publication Date: 2022-05-27
赛尔生命科技(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This serum-free stem cell medium lacks a specialized stem cell medium designed for the technical needs of inducing bone marrow dedifferentiated mesenchymal stem cells with high directional differentiation efficiency and high survival rate to supply bone damage repair rather than superficial wound healing

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 The present invention proposes a supernatant gel stem cell culture medium for bone injury repair. The culture medium includes: IMDM medium with a volume ratio of 1:0.2-0.5:0.01-0.05, suspension of bone marrow mesenchymal stem cells solution and neural induction compound solution.

[0020] As a preferred embodiment, each 500 mL of the IMDM medium consists of the following components: recombinant human insulin 1 mg / mL; recombinant transferrin 2.5 μg / mL; vitamin C 16.1 μg / mL; Protein 1mg / mL; Fibroblast Growth Factor 10ng / mL; Sufficient Human Platelet-Derived Growth Factor 10ng / mL; Growth Promoting Factor IGF-110ng / mL; Transforming Growth Factor TGF-β 10ng / mL; Fibronectin 10ng / mL; Fetuin 1mg / mL; β-cytokinin 10μg / mL; β-mercaptoethanol 1 50μM; ALK inhibitor 1~3mmol / L.

[0021] As a preferred embodiment, the bone marrow mesenchymal stem cell suspension is prepared as follows: 5 g of human bone marrow tissue is cut into pieces, digested with 5 to 10 ml of 0.2% type I...

Embodiment 2

[0023] Example 2 The present invention also proposes a method for preparing a supernatant gel stem cell culture medium for bone injury repair, including the following steps:

[0024] Step SS1: respectively prepare IMDM medium, bone marrow mesenchymal stem cell suspension and neural induction compound solution in a volume ratio of 1:0.2-0.5:0.01-0.05; the bone marrow mesenchymal stem cell suspension in step SS1 The preparation method includes: shredding 5g of human bone marrow tissue, digesting with 5-10ml of 0.2% type II collagenase in a water bath at 15-37°C for 1-3h to obtain a bone marrow mesenchymal stem cell suspension.

[0025] Step SS2: adding the nerve-inducing compound solution prepared in the step SS1 into the IMDM medium prepared in the step SS1 to prepare an induction medium solution;

[0026] Step SS3: add the bone marrow mesenchymal stem cell suspension prepared in the step SS1 to the induction medium solution prepared in the step SS2 and mix it in a water bath a...

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Abstract

The invention discloses a supernatant gel stem cell culture medium for bone injury repair and a preparation method. The culture medium comprises an IMDM culture medium, a bone marrow mesenchymal stem cell suspension and a nerve induction compound solution in a volume ratio of 1: (0.2-0.5): (0.01-0.05). The preparation method comprises the following steps: step SS1, respectively preparing an IMDM culture medium, a bone marrow mesenchymal stem cell suspension and a nerve induction compound solution according to a volume ratio of 1: (0.2-0.5): (0.01-0.05); step SS2, adding the nerve induction compound solution prepared in the step SS1 into the IMDM culture medium prepared in the step SS1 to prepare an induction culture medium solution; and step SS3, adding the bone marrow mesenchymal stem cell suspension prepared in the step SS1 into the induction culture medium solution prepared in the step SS2, and mixing to prepare the supernatant gel stem cell culture medium. According to the method, the technical requirements of obtaining bone marrow dedifferentiated mesenchymal stem cells with high directional differentiation efficiency and high survival rate through induction for bone injury repair instead of surface wound healing are thoroughly met.

Description

technical field [0001] The invention relates to a supernatant gel stem cell culture medium for repairing bone damage and a preparation method thereof, belonging to the technical field of biological materials. Background technique [0002] The wound healing process is a complex process, which involves re-epithelialization of the skin defect, proliferation of granulation tissue, inflammatory response, vascular proliferation, and wound remodeling. Problems in any of these processes will lead to wound healing. obstacle. The Chinese Patent Publication No. CN105797205A in the prior art discloses a stem cell culture supernatant gel, which specifically discloses that the supernatant gel is composed of a stem cell culture supernatant and methyl cellulose, the solvent is water, and the stem cell culture supernatant is mesenchymal. Stem cells are collected and purified by culturing in serum-free stem cell medium. The supernatant gel is used for skin wound repair with remarkable effec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0663C12N2500/25C12N2500/38C12N2501/998C12N2501/115C12N2501/135C12N2501/105C12N2501/15C12N2500/44C12N2501/727C12N2509/00C12N2509/10C12N2501/415
Inventor 王凌鸿谢海涛杨璐
Owner 赛尔生命科技(深圳)有限公司
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