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Cell feeding method for treating heart disease

A technology of heart disease and cells, which is applied in the field of cell delivery for the use of cells to treat heart diseases. It can solve the problems of low cell utilization, invasion specialization, and few treatment cells, so as to repair and improve structure and function, and maintain biological activity. , the effect of a good initial retention rate

Inactive Publication Date: 2015-09-23
SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

⑴ When using the method of intramyocardial injection, the therapeutic cells can directly reach the myocardium of the lesion site, but there are problems such as the small initial retention of the injected cells, the selection of the injection area, the limitation of the treatment site, and the damage of the injection site. solve
This method still has the potential to cause arrhythmia. At the same time, it also has certain requirements for the operation itself and the thickness of the ventricular wall: the invasiveness and professionalism of the operation, the need for color Doppler ultrasound to evaluate the patient's ventricular wall thickness before the operation, and the required ventricular wall thickness. Should be greater than 4mm, otherwise it will cause iatrogenic ventricular wall damage
More importantly, because the transplanted cells directly face the harsh microenvironment induced by the damaged myocardium, including ischemia, inflammation and various pro-apoptotic factors, the long-term survival rate of the cells in vivo is difficult to maintain
⑵Although the injection site of various intracoronary injection methods is close to the heart lesion area, since the therapeutic cells cannot penetrate the vascular endothelial barrier, even if repeated injections are repeated, most of the therapeutic cells will eventually enter the systemic circulation. The initial survival rate of cells is low, the utilization rate of therapeutic cells is low, and there is a risk of inducing complications such as arteriolar embolism and arrhythmia
(3) Intravenous injection, which is rarely used at present, has the advantages of simple operation, less invasiveness, and convenient repetition, but the injection of cells cannot effectively avoid the cell clearance of the reticuloendothelial system in the lung, spleen and other tissues during the systemic circulation , after the first level of elimination, theoretically, there are fewer therapeutic cells that finally reach the heart lesion area, the initial survival rate of cells in the heart lesion tissue is low, the long-term survival rate of cells in the body is low, and the utilization rate of therapeutic cells is low
③Ensure the utilization rate of therapeutic cells. The low cell utilization rate requires a large number of therapeutic cells for heart tissue treatment, resulting in too long culture time in vitro, and it is easy to miss the best treatment period
Therefore, compared with the ideal treatment requirements, the clinical promotion of cell therapy for heart diseases still faces many limiting factors

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Cell therapy of rhesus monkey heart failure model using pericardial cavity delivery method

[0042] a. Cultivation of therapeutic cells;

[0043] In vitro isolation, culture and expansion of rhesus monkey autologous bone marrow mesenchymal stem cells (CD166 positive, CD45 negative) to 1×10 7 After labeling the treated cells with the fluorescent marker DiI (5ug / ml, incubated at 37°C for 30min), the treated cells were digested with trypsin to a single-cell state, centrifuged at 1000 rpm for 3 minutes, and the supernatant was removed for later use;

[0044] b. preparing a suspension of therapeutic cells and culture medium;

[0045] Use 3ml of autologous serum to blend the bone marrow mesenchymal stem cells described in step a to obtain a liquid suspension for use;

[0046] c. Put the suspension into the pericardial cavity;

[0047] Using a rhesus monkey model of chronic heart failure, 3ml of the liquid suspension was inserted into the apex of the pericardial ...

Embodiment 2

[0049] Example 2: Cell therapy of rhesus monkey left anterior descending branch ligation-induced myocardial infarction model by pericardial cavity delivery

[0050] a. Cultivation of therapeutic cells;

[0051] In vitro isolation, culture and expansion of rhesus monkey autologous adipose stem cells (CD44 positive, CD45 negative) to 5×10 6 , rhesus monkey autologous cardiac stem cells (c-kit positive) to 5×10 6 , use the fluorescent marker DiI (5ug / ml, incubate at 37°C for 30min) to label autologous adipose-derived stem cells, use the fluorescent marker DiO (5ug / ml, incubate at 37°C for 30min) to label autologous cardiac stem cells, and routinely trypsinize the treated cells to multiple cells In the clump state, centrifuge at 1000 rpm for 3 minutes, remove the supernatant and set aside.

[0052] b. preparing a suspension of therapeutic cells and culture medium;

[0053] Use 0.5ml Nano Peptide Gel (Product No.: 354250) (0.1%) to blend cardiac stem cells and adipose stem cells...

Embodiment 3

[0057] Example 3: Cell therapy of rhesus monkey right coronary artery occlusion-induced myocardial infarction model using pericardial cavity delivery

[0058] a. Cultivation of therapeutic cells;

[0059] In vitro isolation, expansion, and culture of rhesus monkey autologous cardiac stem cells (c-kit positive) to 1×10 7 , After the treated cells were labeled with fluorescent marker DiI (5ug / ml, incubated at 37°C for 30min), the treated cells were routinely trypsinized to the state of multi-cell clusters, centrifuged at 1000 rpm for 3 minutes, and the supernatant was removed for later use.

[0060] b. preparing a suspension of therapeutic cells and culture medium;

[0061] Autologous cardiac stem cells (c-kit positive) were blended with 0.5ml extracellular matrix gel (400ug / ml) to obtain a colloidal suspension for later use.

[0062] c. Put the suspension into the pericardial cavity;

[0063] A rhesus monkey model of acute myocardial infarction was prepared by balloon injury...

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Abstract

The invention discloses a cell feeding method for treating heart disease. The method comprises the following steps: a, culturing treatment cells; b, preparing a suspension of the treatment cells and a culture medium; and c, placing the suspension into pericardial cavity. The method has advantages as follows: it can be ensured that the treatment cells simultaneously have better initial retention rate, in vivo survival rate and cell utilization rate, bioactivity of the treatment cells after transplantation can be well maintained, and cell treatment effect on heart disease can be further enhanced.

Description

technical field [0001] The invention relates to a cell delivery method for treating heart disease, in particular to a cell delivery method for treating heart disease with cells, which is easy to retain and survive the treatment cells. Background technique [0002] The research level and research results of regenerative medicine are gradually becoming one of the important symbols to measure the level of scientific and technological development and health of a country or region. For patients with heart disease, the regenerative medicine program using cell therapy has incomparable advantages over traditional treatment methods (drug therapy and surgical treatment) in reversing and improving heart function. Various research teams around the world have successively discovered that the use of various therapeutic cells can produce significant therapeutic effects on animal models of heart disease and improve the pumping function of the heart. Its mechanism of action is related to pa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/545A61K35/28A61K35/44A61K35/34A61K35/12A61P9/00
Inventor 王景峰张建华武征
Owner SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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