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A method of transfecting microrna into platelets

A technology for platelets and small plates, applied in the field of transfecting microRNA into platelets, can solve the problems of direct transfection of miRNA technology, such as difficulties and stagnation, and achieve the effect of good clinical application prospects.

Inactive Publication Date: 2016-05-25
LUZHOU MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, due to the lack of nucleus and genomic DNA inside platelets, it is extremely difficult to directly transfect miRNAs in platelets, and the research on platelet miRNA markers has been stagnant. It is urgent to find a method that can directly introduce miRNAs into platelets

Method used

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  • A method of transfecting microrna into platelets
  • A method of transfecting microrna into platelets
  • A method of transfecting microrna into platelets

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1 The method of the present invention transfecting microRNA in platelet

[0024] Take the liposome siPORTTMNeoFXTM and the microRNA to be transfected, make solutions respectively, mix them, and place them for 10 minutes at a temperature of 15°C and an oscillation frequency of 50Hz to obtain a mixed solution. The concentration of microRNA in the mixed solution is 0.7mol / L , the concentration of liposomes is 5% (v / v);

[0025] Under the conditions of a temperature of 22° C. and an oscillation frequency of 60 Hz, co-culture the aforementioned mixture with the platelets for 12 hours, centrifuge and collect the platelets.

Embodiment 2

[0026] Embodiment 2 The present invention transfects the method for microRNA in platelet

[0027] Take liposome Lipofectamine2000 and the microRNA to be transfected, make solutions respectively, mix them, and place them for 30 minutes at a temperature of 25°C and an oscillation frequency of 70Hz to obtain a mixed solution. The concentration of microRNA in the mixed solution is 0.8mol / L , the concentration of liposomes is 7% (v / v);

[0028] Under the condition that the temperature is 22° C. and the oscillation frequency is 60 Hz, the aforementioned mixture is co-cultured with the platelets for 36 hours, centrifuged, and the platelets are collected.

Embodiment 3

[0029] Embodiment 3 The present invention transfects the method for microRNA in platelet

[0030] Take liposome Lipofectamine2000 and the microRNA to be transfected, make solutions respectively, mix them, and place them for 20 minutes at a temperature of 22°C and an oscillation frequency of 60 Hz to obtain a mixed solution. The concentration of microRNA in the mixed solution is 0.74mol / L , the concentration of liposomes is 5.9% (v / v);

[0031] Under the condition that the temperature is 22° C. and the oscillation frequency is 60 Hz, the aforementioned mixture is co-cultured with the platelets for 36 hours, centrifuged, and the platelets are collected.

[0032] The method of the present invention can be effectively transferred into platelets by using the method of the experimental example as follows:

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Abstract

The invention discloses a method for transfecting microRNA (Ribonucleic Acid) to blood platelets. The method comprises the following steps: a) uniformly mixing microRNA to be transfected with lipidosomes to obtain a mixed solution; b) co-culturing the mixed solution obtained in the step a with the blood platelets at a temperature of 22 DEG C and an oscillation frequency of 60 Hz for 12-36 hours, centrifuging, and collecting the blood platelets. Through the adoption of the method, the microRNA can be effectively transfected to the blood platelets, and the defects in the prior art are overcome.

Description

technical field [0001] The invention relates to a method for transfecting microRNA into platelets. Background technique [0002] Platelets are one of the key components of thrombus formation in vivo, and play an important role in various pathophysiological processes such as cardiovascular diseases (proliferation, thrombosis, and thromboembolism), inflammation, and some tumors. microRNA (miRNA) is a class of non-coding single-stranded RNA molecules encoded by endogenous genes with a length of 20-24 nucleotides, which participate in post-transcriptional gene expression regulation in animals and plants. [0003] Studies have shown that platelets contain a large number of miRNAs, and some platelet miRNAs related to fibrinolysis and coagulation in vivo may be potential molecular markers of thrombus formation in vivo. [0004] However, due to the lack of nucleus and genomic DNA inside platelets, it is extremely difficult to directly transfect miRNAs in platelets, making the study...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/87C12N5/078
Inventor 罗茂吴剑波李蓉
Owner LUZHOU MEDICAL COLLEGE