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Method for enhancing expression of P-selectin in vascular endothelial cells

A technology of vascular endothelium and P-selectin, which is applied in the fields of biochemical equipment and methods, measurement/inspection of microorganisms, electric/wave energy processing enzymes, etc., can solve the problems of lack of technology, shortage, and ineffective homing of stem cells, etc. To achieve the effect of simple operation and remarkable effect

Pending Publication Date: 2019-01-29
华子昂 +2
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, many technical aspects of stem cell transplantation have not been overcome, which obviously affects the success rate of treatment. The typical problem is that stem cells cannot effectively home to the damaged part.
Many research results have shown that: P-selectin (P-Selectin) of damaged cells is one of the key factors for inducing stem cell homing. Without P-selectin, stem cells cannot locate and adhere to vascular endothelial cells.
To enhance the enrichment of stem cells at the target site, the expression of P-selectin must be effectively enhanced, but there is still a lack of corresponding technology for how to simply and effectively regulate the expression of P-selectin gene in target cells

Method used

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  • Method for enhancing expression of P-selectin in vascular endothelial cells

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Comparison scheme
Effect test

Embodiment 1

[0017] 1) Take out the cryopreservation tube of primary human cardiac microvascular endothelial cells (HCMEC) from liquid nitrogen, melt it quickly in a 37°C water bath, transfer it to a culture bottle coated with L-polylysine, and put a certain amount of endothelial The cell culture medium was cultivated in a 37°C, 5% carbon dioxide incubator, the medium was changed every 2-3 days, and the cells were passaged once every 4-5 days. The cells were digested with trypsin, and the fifth generation cells were used in the experiment.

[0018] 2) After digesting and centrifuging the 5th generation HCMEC cells, resuspend them with 10% complete culture medium and transplant them into a culture dish; sterilize the shock wave probe, set the parameters to 2.5bar, 10Hz, and use shock waves to act on the cells 200 times , the control group was the fifth generation HCMEC cells without shock wave treatment, and the shock wave probe was selected from the Swiss EMS company.

[0019] 3) The cells...

Embodiment 2

[0025] 1) Take out the cryopreservation tube of primary human cardiac microvascular endothelial cells (HCMEC) from liquid nitrogen, melt it quickly in a 37°C water bath, transfer it to a culture bottle coated with L-polylysine, and put a certain amount of endothelial The cell culture medium was cultured in a 37°C, 5% carbon dioxide incubator, the medium was changed every 2-3 days, and the cells were passaged once every 4-5 days. The cells were digested with trypsin; the fifth passage cells were used in the experiment.

[0026] 2) After digesting and centrifuging the 5th generation HCMEC cells, resuspend them with 10% complete culture medium and transplant them into a petri dish; sterilize the shock wave probe, set the parameters to 1.5bar, 10Hz, and use shock waves to act on the cells 200 times , the control group was the fifth passage HCMEC cells without shock wave treatment.

[0027] 3) The cells subjected to the shock wave treatment were cultured for 60 hours for RNA isolat...

Embodiment 3

[0030] 1) Take out the cryopreservation tube of primary human cardiac microvascular endothelial cells (HCMEC) from liquid nitrogen, melt it quickly in a 37°C water bath, transfer it to a culture bottle coated with L-polylysine, and put a certain amount of endothelial The cell culture medium was cultured in a 37°C, 5% carbon dioxide incubator, the medium was changed every 2-3 days, and the cells were passaged once every 4-5 days. The cells were digested with trypsin; the fifth passage cells were used in the experiment.

[0031] 2) After digesting and centrifuging the 5th generation HCMEC cells, resuspend them with 10% complete culture medium and transplant them into a petri dish; sterilize the shock wave probe, set the parameters to 2.0bar, 10Hz, and use shock waves to act on the cells 200 times , the control group was the fifth passage HCMEC cells without shock wave treatment.

[0032] 3) The cells subjected to the shock wave treatment were cultured for 60 hours for RNA isolat...

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Abstract

The invention provides a method for enhancing the expression of P-selectin in vascular endothelial cells. The method comprises the following steps: (1) setting the action mode and action parameters, including pressure, power frequency and action frequency, of shock wave equipment; (2) preparing vascular endothelial cells for subculture or vascular endothelial cells in experimental mice, performingcentrifugation, carrying out resuspending by using a 10% complete culture medium, transferring the resuspended cells to a Petri dish, and then subjecting the cells to shock wave treatment via a probe; and (3) after completion of the shock wave treatment, continuing to culture for 60 hours, then separating the vascular endothelial cells, and detecting the genetic expression quantity of P-selectinby using a real-time fluorescent quantitative PCR method, with a GAPDH gene used as a reference internal standard. According to the invention, the expression of P-selectin in the vascular endothelialcells is enhanced by shock waves without other drugs or chemical stimulation, so the method is simple in operation and remarkable in effect, especially has absolute advantages in the expression of P-selectin in in-vivo vascular endothelial cells, and is a basis for the future development of stem cell homing and stem cell-related technologies.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for enhancing the expression of P-selectin in vascular endothelial cells. Background technique [0002] Stem cells are called "universal cells" in the medical field, which can differentiate into various functional cells or tissues and organs. Stem cell transplantation therapy is to transplant healthy stem cells into the patient's body to repair or replace damaged cells or tissues, so as to achieve the purpose of cure. Stem cell transplantation has a wide range of treatments, and can generally treat nervous system diseases, immune system diseases, and other medical and surgical diseases. However, many technical aspects of stem cell transplantation have not been overcome, which obviously affects the success rate of treatment. A typical problem is that stem cells cannot effectively home to the damaged site. Many research results have shown that: P-selectin (P-Selectin) of dam...

Claims

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Application Information

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IPC IPC(8): C12P21/00C12N13/00C12Q1/6851
CPCC12N13/00C12P21/00C12Q1/6851C12Q2531/113C12Q2545/101
Inventor 华子昂
Owner 华子昂
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