Culturing method of human peripheral blood vessel endothelial ancestry cell

A technology of endothelial progenitor cells and human peripheral blood, which is applied in the field of culturing human peripheral blood vascular endothelial progenitor cells, can solve the problems of poor repeatability, MNC loss, and high experimental conditions

Inactive Publication Date: 2003-04-16
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The traditional culture method is cumbersome. In the process of sorting MNC with CD34+ antibody-coated immunomagnetic beads, the loss of MNC CD34+ is inevitable, resulting in a low cell yield.
In addition, foreign traditional methods require high experimental conditions, so the repeatability of the method is poor

Method used

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  • Culturing method of human peripheral blood vessel endothelial ancestry cell
  • Culturing method of human peripheral blood vessel endothelial ancestry cell
  • Culturing method of human peripheral blood vessel endothelial ancestry cell

Examples

Experimental program
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Embodiment 1

[0029] 1. Isolation of MNC from human peripheral blood: Take 10-15ml of human femoral artery anticoagulant blood, dilute it with D-Hanks balanced salt solution 1:1, spread it on the Ficoll separation medium along the tube wall, and centrifuge at 2000rpm×30min density gradient to remove white The cloud layer was sucked out, and then washed twice with D-Hanks balanced salt solution 1000rpm×10min, and finally the cells were resuspended in culture medium and counted with trypan blue.

[0030] 2. Cultivate MNC in vitro: the culture medium is M-199+20%FBS+heparin 100U / ml+0.1mg / ml penicillin+0.1mg / ml streptomycin; the growth factor is bovine pituitary extract with a final concentration of 9 μg / ml Added to the culture medium; the cells were pressed by 10 5 / cm 2 Sow in culture flasks or slides coated with 2% gelatin at a gelatin density of 5 μl / cm 2 ; Cells were placed at 37°C, 5% CO 2 In an incubator, change the medium in half every 3-4 days.

[0031] 3. Identification of the cha...

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Abstract

A method for culturing human peripheral blood vascular endothelial archeocyte features that the MNC cell is directly used for culturing in vitro to obtain EPCs. Its step are adding a growth factor tothe culture liquid, implanting in culture bottle or on glass sheet by a certain density, and conventional culture. Its advantages are simple process, high output rate of EPCs, and low cost.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a culture method for human peripheral blood vascular endothelial progenitor cells. Background technique [0002] Artificially stimulating angiogenesis is a new approach to treating ischemic diseases. Angiogenesis includes two concepts: embryonic angiogenesis (vasculogenesis, VG) and postnatal angiogenesis (angiogenesis, AG). In the traditional sense, VG refers to the differentiation of endothelial progenitor cells (endothelial progenitor cells, EPCs) or angioblasts (angioblasts) into endothelial cells in the absence of a vascular system, which sprouts to form a vascular network, and then forms blood vessels. This process usually begins with hematopoietic stem cells (HSCs) at the center and EPCs in the surrounding blood islands. AG in the traditional sense means that in adult blood vessels, the existing mature endothelial cells break through the wall matrix to mi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078C12Q1/02C12Q1/68
Inventor 孙宝贵严轶文戴秋艳
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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