Umbilical cord mesenchymal stem cells (MSCs) and culture method and application thereof

A kind of stem cell and culture method technology, applied in the field of umbilical cord mesenchymal stem cell MSCs and its culture, can solve the problems of hindering the clinical treatment of stem cells, cell mass embolism, stem cell adhesion, etc., to improve the ability of cell proliferation and clone formation, Enhances adaptability and reduces the effect of clumping

Active Publication Date: 2019-08-23
广东新南方干细胞再生医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, MSCs have the characteristics of adherent growth, and MSCs often form clusters in vitro culture. Therefore, after intravenous infusion into the human body, stem cells will adhere, aggregate into clusters, and cell clusters embolize.
[0006] For the above reasons, MSCs cultured in vitro will cause adverse reactions in the treatment of diseases, affect the curative effect, and hinder the clinical treatment of stem cells.

Method used

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  • Umbilical cord mesenchymal stem cells (MSCs) and culture method and application thereof
  • Umbilical cord mesenchymal stem cells (MSCs) and culture method and application thereof
  • Umbilical cord mesenchymal stem cells (MSCs) and culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] A kind of MSC stem cell obtained by cultivating by the following method:

[0053] 1. Donor Screening

[0054] 1. Donors should draw blood within one month before sample collection for detection of severe infectious diseases, including but not limited to: HIV (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), Treponema pallidum (TP), giant cells Virus (CMV), Epstein-Barr virus, phagocytic T-cell virus (HLTV), any one of the seven infectious disease tests showing that the donors carry pathogens (including Treponema pallidum previous infection) are excluded;

[0055] 2. The donor draws blood within one month before sample collection for DNA testing of serious genetic diseases, including but not limited to: achondroplasia, congenital deafness, vitamin D-resistant rickets, phenylketonuria, hemophilia, progressive Muscular dystrophy, glucose 6-phosphate dehydrogenase (G-6-PD) deficiency (faba bean disease), etc.; tested positive, are excluded);

[0056] 3. The donor ...

Embodiment 2

[0096] A kind of MSCs stem cell injection, obtained by the following method:

[0097] 1. Preparation:

[0098] 1.1 Turn on the water bath for rewarming to stabilize the water temperature at 37°C;

[0099] 1.2 The biosafety cabinet is sterilized and ventilated by ultraviolet light 30 minutes in advance;

[0100] 1.3 Prepare normal saline according to the number of recovered cells and add it to the centrifuge tube. The volume of normal saline should be at least 10 times the volume of the whole cryopreservation solution;

[0101] 2. Information check:

[0102] Take out the frozen cells from the liquid nitrogen tank according to the predetermined requirements, and carefully check key information such as cell type, cell code, cell number, and cell generation to prevent errors.

[0103] 3. Immediately put the frozen cells taken out of the liquid nitrogen tank into the rewarmed water bath to resuscitate for 2 minutes until the frozen storage solution is completely melted, keep sha...

Embodiment 3

[0118] An application of MSCs stem cell injection in the treatment of myocardial infarction.

[0119] The umbilical cord mesenchymal stem cell MSCs preparation of the present invention has a wide range of applications. Currently, we have formulated the MSCs of Example 1 into a composite preparation as shown in Example 2 and applied it to the treatment of patients with myocardial infarction to observe the curative effect.

[0120] 1. Observe the morphology of peripheral blood cells under a microscope.

[0121] 1. Method

[0122] The peripheral blood of subjects in the control group (the MSC stem cell culture was not added Ligustrazine hydrochloride and Shenmai injection) and the administration group (composite preparation of Example 2) was observed under a microscope.

[0123] 2. Results

[0124] The result is as Figure 10-11 as shown, Figure 10 Peripheral blood condition of the subjects in the control group, Figure 11 Peripheral blood condition of subjects in the admin...

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Abstract

The invention relates to umbilical cord mesenchymal stem cells (MSCs) and a culture method and application thereof, and belongs to the technical field of biological medicine. The culture method includes MSCs culture steps and phenotypic examination. The MSCs culture steps include primary culture and sub-culturing; during primary culture, Huatong gum tissue of the umbilical cord is obtained and cultured in a serum-free culture medium under hypoxic conditions; during sub-culturing, P1 primary cells are collected, a single cell suspension is prepared, cell precipitates are obtained after centrifuging, the serum-free culture medium is added into the cell precipitates, culturing is conducted under hypoxic conditions until next generations come out, continuous culturing is performed to obtain P2-P3 generations, ligustrazine hydrochloride and Shenmai injections are added in each sub-culturing stage, when the cells grow to a predetermined fusion degree, digestion is performed and the cells arecollected until the P6 generations come out. During phenotypic examination, the collected cells are subjected to phenotypic examination for use. An MSCs preparation obtained through culturing in themethod reduces the easy-to-aggregation characteristics of the stem cells, the occurrence of cell adhesion, erythrocyte rouleaux formation and cell mass embolism after intravenous infusion into the human body is thus avoided, and the stem cell (MSCs) technology can be better applied to clinical practice.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an umbilical cord mesenchymal stem cell MSCs and a culture method and application thereof. Background technique [0002] According to the forecast of the World Health Organization, by 2020, non-communicable diseases will account for 79% of the causes of death in my country, and cardiovascular diseases will take the first place. Since cardiomyocytes are terminally differentiated cells, necrotic cells cannot be regenerated and can only be replaced by scar formation, resulting in loss of myocardial contractile function, refractory heart failure and death. According to the case survey of 50 tertiary hospitals in my country, although the hospitalization rate of heart failure accounts for 20% of cardiovascular diseases in the same period, the mortality rate accounts for 40%. The five-year survival rate is similar to that of malignant tumors. Clinical drugs, interventional and sur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K35/51A61K9/08A61P9/10A61K31/195A61K31/198A61K31/727
CPCC12N5/0668A61K35/51A61K31/198A61K31/195A61K31/727A61K9/0019A61K9/08A61P9/10A61K2300/00A61K35/28A61K2035/124C12N2500/02C12N2500/76C12N2500/99C12N2501/999C12N5/0665C12N2500/30C12N2500/90
Inventor 黄建文
Owner 广东新南方干细胞再生医学科技有限公司
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