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Stem cell preparation for treating ischemic cardiomyopathy and preparation method of stem cell preparation

A technology of ischemic cardiomyopathy and stem cell preparation, applied in the field of stem cell biotechnology and cell transplantation, can solve the problems of limitations in stem cell growth and differentiation ability, difficulty in supplying stem cells, etc., achieve excellent heart repair ability, improve capillary Density, low immune rejection effect

Inactive Publication Date: 2014-06-18
奥思达干细胞有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of autologous stem cells has relatively good safety, but because it comes from the patient itself, the growth and differentiation ability of stem cells has certain limitations, especially some patients may find it difficult to provide their own stem cells due to their own diseases

Method used

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  • Stem cell preparation for treating ischemic cardiomyopathy and preparation method of stem cell preparation
  • Stem cell preparation for treating ischemic cardiomyopathy and preparation method of stem cell preparation
  • Stem cell preparation for treating ischemic cardiomyopathy and preparation method of stem cell preparation

Examples

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Effect test

Embodiment 1

[0042] Example 1: Experimental conditions for preparing stem cells: In order to ensure the standardization and standardization of cell collection, separation, culture, and cryopreservation in vitro, to obtain cells that meet the clinical application level, and to ensure that cells can treat diseases in a safe and effective manner, the following guidelines should be followed: Relevant regulations at home and abroad, as well as the requirements of China's Good Manufacturing Practice (GMP), established a series of standard operating procedures (Standard Operating Procedure, SOP) for in vitro cell operations.

Embodiment 2

[0043] Example 2: The preparation method of placenta and umbilical cord stem cells, i.e. mesenchymal stem cells, is: the placenta and umbilical cord tissue from which blood stains have been removed are mechanically shredded into 1mm 3 The small pieces were infiltrated with collagenase Ⅰ with a concentration of 1mg / ml, digested at 37°C for 3 hours, and gently shaken once every 15 minutes; the cells were collected by filtering through a 100-mesh sieve; the cells were washed 3 times with D-Hank's balanced salt solution, Centrifuge at 200g for 8 minutes; resuspend the cells in high-glucose DMEM medium containing 10% fetal bovine serum, and adjust the cell density to 4.8×10 3 ~1×10 4 / cm 2 , inoculated in T75 culture flasks, as primary cells, at 37°C, with a volume fraction of 5% CO 2 Cultivate in the incubator, replenish the liquid after 48 hours, and change the liquid after 72 hours; when the cells grow to 80% confluent, digest with a mixture of 0.25% trypsin and 0.02% EDTA, ob...

Embodiment 3

[0047] Example 3: The preparation method of myocardial stem cells is: take the third to fifth generation partial placenta and umbilical cord tissue stem cell suspension, and the cell concentration is 5×10 5 / dish was inoculated in 100mm petri dish, and after 24 hours of adherence, an induction system was added, using a low-sugar DMEM culture system with 10% fetal bovine serum, which contained 10 μmol / L of 5-azacytidine, and each dish contained 8 ml of induction system. After 24 hours of induction, the induction system was absorbed, washed with D-Hank's balanced salt solution, and then replaced with 10% fetal bovine serum low-sugar DMEM culture line to continue culturing for 2 weeks. Then it was expanded for three generations and collected for use; the growth state of cardiac stem cells after successful induction was also long spindle cells, such as figure 1 Shown in B.

[0048] Determination of the growth activity of cardiac stem cells: collect cardiac stem cells induced and ...

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Abstract

The invention discloses a stem cell preparation for treating ischemic cardiomyopathy and a preparation method of the stem cell preparation. The preparation consists of mesenchymal stem cells, cardiac stem cells and vascular endothelial stem cells, and a solvent is platelet-rich plasma lysate of a patient. The stem cells express a corresponding specific identifier and do not express a corresponding non-specific identifier at the same time. The preparation method comprises the following steps: (1), preparing mesenchymal stem cells between a placenta and an umbilical cord; (2), applying different induction culture systems to induce the mesenchymal stem cells in the step (1) to be cardiac stem cells and vascular endothelial stem cells; (3), preparing the solvent which is the platelet-rich plasma lysate; and (4), uniformly mixing and preparing the stem cell preparation. The stem cell preparation disclosed by the invention is rich in resource, easy to prepare, convenient to collect, and free of pain while being applied; moreover, the stem cell preparation is low in immunological rejection after being transplanted. The stem cell preparation comprises a plurality of stem cells, so that advantages of combined effect of a plurality of cells can be brought into play; the solvent is easily available, and can directly act on a patient; besides, transplantation effect is enhanced.

Description

technical field [0001] The invention relates to a stem cell preparation for treatment and a preparation method thereof, in particular to a stem cell preparation for ischemic cardiomyopathy transplantation and a preparation method thereof, and belongs to the field of stem cell biotechnology and cell transplantation. Background technique [0002] At present, the incidence and mortality of cardiovascular diseases rank first in the world, and ischemic cardiomyopathy (ICM) accounts for more than 50% of cardiovascular diseases, causing about 17 million deaths per year. death, and the incidence of ischemic cardiomyopathy in my country is still increasing year by year. Ischemic cardiomyopathy belongs to a special type or late stage of coronary heart disease, which refers to the myocardial lesion mainly caused by diffuse fibrosis caused by long-term myocardial ischemia due to coronary atherosclerosis, also known as myocardial sclerosis or cardiac muscle fibrosis. The main cause of t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/50A61K35/44A61P9/10C12N5/0775C12N5/073A61K35/34A61K35/14A61K35/16A61K35/28
Inventor 周萱
Owner 奥思达干细胞有限公司
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