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Application of mesenchymal stem cell modified by miR-21 antisense nucleotide

A stem cell and product technology, applied in the field of adult stem cells, can solve the problems of inability to produce obvious and durable therapeutic effects, loss of biological activity, and uneven implantation efficiency in cases, and achieve significant immunity, stable biological performance, and product harmless effect

Inactive Publication Date: 2015-11-25
CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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Problems solved by technology

The problem it faces is that the implantation efficiency is not uniform, and some cases cannot produce obvious and lasting therapeutic effects
In addition, with the extension of in vitro culture time, MSCs will lose part of their biological activity, reducing the application efficiency.

Method used

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  • Application of mesenchymal stem cell modified by miR-21 antisense nucleotide
  • Application of mesenchymal stem cell modified by miR-21 antisense nucleotide
  • Application of mesenchymal stem cell modified by miR-21 antisense nucleotide

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Experimental program
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Embodiment Construction

[0029] cell culture

[0030] Experimental materials Healthy human bone marrow comes from patients with trauma or orthognathic surgery who need fibula transplantation, healthy human periodontal tissue comes from non-infected premolars or third molars due to orthodontic needs, and healthy human deciduous tooth pulp tissue comes from clinic due to retention Baby teeth that need to be extracted.

[0031] 1. Culture of human odontogenic MSCs

[0032] Human Dental Pulp Stem Cell Culture

[0033] Freshly extracted teeth were immediately placed into centrifuge tubes filled with sterile PBS and antibiotics, and dental pulp stem cells were isolated and cultured within 12 hours. Wrap the hard tissue of the tooth with sterile gauze, and beat it with a hammer soaked in alcohol to obtain the pulp tissue, wash it repeatedly with PBS, cut it into pieces as much as possible, and put it in a solution containing 3 mg / ml type I collagenase and 4 mg / ml Dispase , digested in a water bath at 37°C...

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Abstract

The research group selects the optimum method for preparing a product of the invention, namely, enabling 100nM miR-21inhibitor transfected by lipo 2000 RNAimax to enter the stem cell. The stem cell product prepared by the method is enhanced in the capacity of secreting transforming growth factor TPF-beta 1, thus the capacity of inducing the T cell to differentiate towards regulatory T cells after co-culture of the cell and the T cell is enhanced, namely, the immunoregulation capacity of the stem cell is enhanced. In an in-vivo experiment, a medicine is adopted for inducing a mouse for establishing an acute enteritis model, the stem cell and a control cell which are transfected by miR-21 inhibitor are transfused into the body of the mouse through caudal vein by using the dosage of 1X10<6>, the result shows that the clinical symptoms and the immune indices of the mouse with stem cell group transfected by miR-21 inhibitor transfused are superior to those of the control cell group with transfusion, namely, the stem cell transfected by miR-21 inhibitor can be used for more effectively treating mouse enteritis. All in all, the stem cell with strong and stable effect is prepared, and experimental basis is provided for improving the application efficiency of the stem cell in clinic.

Description

technical field [0001] The invention relates to the field of adult stem cells. The expression level of endogenous miR-21 in mesenchymal stem cells (mesenchymalstemcells, MSCs) is silenced by specific antisense nucleotide sequences to enhance the immune regulation ability of MSCs, and the modified MSCs are studied use. Background technique [0002] In the traditional concept, MSCs such as bone marrow mesenchymal stem cells were first isolated and proved to be able to differentiate into different tissue cells such as bone, cartilage, fat, islets, liver and nerves under different induction environments, and are currently the most commonly used seed cells for the treatment of bone diseases , but with the prolongation of the in vitro culture time, the osteogenic ability of the implanted body decreased significantly, and the multidirectional differentiation ability was gradually lost. In addition, studies have shown that bone marrow mesenchymal stem cells can play an important ro...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K48/00A61P37/02
Inventor 王松灵吴婷婷张春梅
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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