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Preparation method and application of targeting mesenchymal stem cell (MSC) drug delivery system

A mesenchymal stem cell and delivery system technology, applied in the field of preparation of targeted mesenchymal stem cell drug delivery system, can solve the problems of unfavorable inflammation and fibrosis repair, large individual differences, unstable curative effect, etc., with excellent ability and no ethics problems, avoid blood flow washing or natural shedding effect

Inactive Publication Date: 2020-02-11
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although most experiments have shown that MSCs have a repairing effect on tissue damage, the curative effect is not stable and individual differences are large; some studies have shown that MSCs transplantation has no effect on the repair of tissue function and tissue structure, and even has a negative impact
The reason is that, on the one hand, transplanted MSCs can only effectively play a therapeutic and repairing role when they colonize / homing to the lesion, but in vivo studies have found that only a small amount of transplanted cells are enriched to the injury or lesion; secondly, the transplanted MSCs Colonize to the tissue injury site, some differentiate into tissue macrophages, and tissue macrophages can promote the activation, proliferation, migration, and survival of tissue astrocytes, which are not conducive to the repair of inflammation and fibrosis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The present invention provides a technical solution: a preparation method and application of a drug delivery system targeting mesenchymal stem cells, the method comprising the following steps:

[0021] S1: Collect stem cells, collect mesenchymal stem cells and inoculate them in a 75cm2 culture flask, 10% FBS+L-DMEM, 37°C, saturated humidity, 5% CO2 incubator;

[0022] S2: Detect cells, take the 1st generation of subcultured cells, make a single cell suspension, use 0.4% trypan blue as the staining agent, count live cells and dead cells with a cell counting board, a total of 3 times, take the average value of the 3 times, and calculate cell viability;

[0023] S3: Making a transfection mixture, diluting the miR-26b mimic and the transfection reagent with L-DMEM respectively, mixing them, and standing at room temperature to obtain a transfection mixture;

[0024] S4: pretreatment of mesenchymal stem cells, the mesenchymal stem cells cultured to the 4th passage and reachi...

Embodiment 2

[0029] The present invention provides a technical solution: a preparation method and application of a drug delivery system targeting mesenchymal stem cells, the method comprising the following steps:

[0030] S1: Collect stem cells, collect mesenchymal stem cells and inoculate them in a 75cm2 culture flask, 10% FBS+L-DMEM, 37°C, saturated humidity, 5% CO2 incubator;

[0031] S2: Detect cells, take 3 passaged cells, make a single cell suspension, use 0.4% trypan blue as the staining agent, count live cells and dead cells with a cell counting board, a total of 3 times, take the average of 3 times, and calculate cell viability;

[0032] S3: Making a transfection mixture, diluting the miR-26b mimic and the transfection reagent with L-DMEM respectively, mixing them, and standing at room temperature to obtain a transfection mixture;

[0033] S4: pretreatment of mesenchymal stem cells, the mesenchymal stem cells cultured to the 5th generation and reaching a confluence of 85% were wa...

Embodiment 3

[0038] The present invention provides a technical solution: a preparation method and application of a drug delivery system targeting mesenchymal stem cells, the method comprising the following steps:

[0039] S1: Collect stem cells, collect mesenchymal stem cells and inoculate them in a 75cm2 culture flask, 10% FBS+L-DMEM, 37°C, saturated humidity, 5% CO2 incubator;

[0040] S2: Detect cells, take 4 passaged cells, make single cell suspension, use 0.4% trypan blue as staining agent, count living cells and dead cells with a cell counting board, a total of 3 times, take the average value of 3 times, and calculate cell viability;

[0041] S3: Making a transfection mixture, diluting the miR-26b mimic and the transfection reagent with L-DMEM respectively, mixing them, and standing at room temperature to obtain a transfection mixture;

[0042] S4: pretreatment of mesenchymal stem cells, the mesenchymal stem cells cultured to the 6th passage with a confluence of 88% were washed with...

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Abstract

The invention discloses a preparation method and application of a targeting mesenchymal stem cell (MSC) drug delivery system. The preparation method comprises the steps as follows: S1, stem cell collection; S2, cell detection; S3, production of a transfection mixed solution; S4, pretreatment of MSCs; and S6, alkaline culture. The MSCs are widely sourced, can be xenogenous as well as autogenous andhave sufficient sources, and the cells have excellent capacity in proliferation, survival, differentiation, migration, synthesis, factor secretion and other aspects, are free of ethical problems andhave low immunogenicity and broad application prospects. The disclosed tissue damage targeting MSCs can be effectively planted at tissue damage parts, blood washing or natural falling can be avoided,the homing cell quantity can be increased, besides, low expression of smad protein can be avoided, and the fibrosis promotion risk due to transplantation of the MSCs can be avoided.

Description

technical field [0001] The invention relates to the field of drug delivery, in particular to a preparation method and application of a drug delivery system targeting mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells are a kind of multipotent stem cells, which have all the common characteristics of stem cells, that is, self-renewal and multidirectional differentiation capabilities. At present, it is also the most clinically used, and combined with hematopoietic stem cells can improve the success rate of transplantation and accelerate hematopoietic reconstruction. After the patient receives high-dose chemotherapy, the infusion of mesenchymal stem cells and hematopoietic stem cells can significantly accelerate the recovery time of the patient's blood cells, and it is safe and has no adverse reactions. Mesenchymal stem cells exist not only in bone marrow, but also in skeletal muscle, periosteum and trabecular bone. Because it differentiates a wide ra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/113
CPCC12N5/0605C12N5/0663C12N5/0664C12N5/0665C12N5/0667C12N5/0668C12N15/113C12N2310/141C12N2501/115C12N2510/00
Inventor 段莉王大平徐晓李兴福熊建义欧阳侃
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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