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Separation, screening, culture and functional identification method of human aortic vascular wall stem cells expressing c-Kit

A technology for aorta and blood vessel wall, which is applied in cell dissociation methods, vascular endothelial cells, cell culture active agents, etc., can solve the problems of low purity of stem cells and molecular screening of stem cell markers, and achieve improved purity and cell activity, good Effects of In Vitro Proliferation and Migration Ability

Active Publication Date: 2018-11-06
CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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Problems solved by technology

Although some literatures have reported that cells isolated from normal human aortic vessels express mesenchymal stem cell marker molecules, the screening of stem cell marker molecules was not carried out during the isolation process, resulting in the fact that the purity of the stem cells obtained in the end is not high

Method used

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  • Separation, screening, culture and functional identification method of human aortic vascular wall stem cells expressing c-Kit
  • Separation, screening, culture and functional identification method of human aortic vascular wall stem cells expressing c-Kit
  • Separation, screening, culture and functional identification method of human aortic vascular wall stem cells expressing c-Kit

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Embodiment 1

[0048] Example 1. Isolation, screening and cultivation of human aortic vessel wall stem cells

[0049] 1. Isolation and screening of human aortic vessel wall stem cells

[0050] 1. Isolation of human aortic vessel wall stem cells

[0051] 1) Separation of adventitia

[0052]Aortic tissue from normal organ donation donors or patients undergoing aortic replacement (patients, donors or donor family members informed), the area is about 2×5cm 2 , immediately put into 20ml ice-cold sterile tissue preservation solution (containing 1mg EDTA-Na 2 and 3% FBS in sterile PBS). Use sterile tweezers to remove the vascular tissue from the tissue preservation solution in an ultra-clean bench, and use 1 mg EDTA-Na 2 Rinse the vascular tissue with sterile PBS solution several times to remove residual blood, until the rinsing solution is clear and no blood is visible to the naked eye. Use two sterile tweezers to gently separate the vascular adventitia from the media, retain the adventitial ...

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Abstract

The invention discloses a separation, screening, culture and functional identification method of human aortic vascular wall stem cells expressing c-Kit and provides a kit for obtaining human aortic vascular wall stem cells. The kit contains a substance bound with c-Kit positive cells. In the kit, the substance bound with the c-Kit positive cells includes a c-Kit antibody, and / or the c-Kit antibodyis specifically a c-Kit immunomagnetic bead antibody. Tunica adventitia vasorum is separated from human aortic tissue and is digested with collagenase, a cell suspension is obtained through filtration of a cell strainer, c-Kit positive vascular wall stem cells are obtained through screening of the c-Kit immunomagnetic bead antibody, the stem cells have good in-vitro proliferation and transfer ability, has the differentiation potential of mesenchymal stem cells (can be differentiated into fat cells, osteoblasts and cartilage cells) and also has the tissue specificity differentiation capacity of differentiating vascular wall stem cells into vascular smooth muscle cells, and apoptosis occurs rarely.

Description

technical field [0001] The invention belongs to the technical field of stem cells and tissue engineering, and in particular relates to a method for separating, screening, cultivating and functionally identifying human aortic vessel wall stem cells expressing c-Kit. Background technique [0002] Adult stem cells are a type of undifferentiated cells that exist in differentiated tissues in adult animals or humans. These cells have the ability to continuously proliferate and self-renew, and can differentiate into specific types of cells in the tissue in which they are located, thereby replenishing the tissue in time. Damaged mature cells maintain the dynamic balance of tissue and organ growth and aging, damage and repair. In recent years, a large number of studies have shown that a group of adult stem cells resides in the adventitia of arteries and veins in mice and humans. Under normal physiological conditions, they maintain vascular homeostasis and replenish and update aging a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0692C12N2509/00C12N2501/235C12N2501/115C12N2500/44
Inventor 于宝琪曲爱娟
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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