Unlock instant, AI-driven research and patent intelligence for your innovation.

A method for extracting, separating and culturing fetal mouse endothelial progenitor cells

An endothelial progenitor cell, isolation and culture technology, applied in cell dissociation methods, vascular endothelial cells, animal cells, etc., can solve the problems of a small number of endothelial progenitor cells, expansion efficiency and slow cell proliferation, and achieve an easy separation method. , Great application prospects and value, the effect of abnormal proliferation ability

Inactive Publication Date: 2020-03-10
CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The key to the clinical application of endothelial progenitor cells is to explore their ideal tissue source, but there are great differences in the methods of recruiting, isolating and culturing endothelial progenitor cells, and the number of endothelial progenitor cells obtained is small, the expansion efficiency and cell proliferation are slow

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A method for extracting, separating and culturing fetal mouse endothelial progenitor cells
  • A method for extracting, separating and culturing fetal mouse endothelial progenitor cells
  • A method for extracting, separating and culturing fetal mouse endothelial progenitor cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Extraction, isolation and culture of endothelial progenitor cells from fetal rat lung

[0059] The implementation steps are as follows:

[0060] (1) Purchasing healthy adult clean SD rats (female / male) at the Experimental Animal Center of Chongqing Medical University. When the rats are in estrus, place 2 female rats and 1 male rat in the same rat box overnight. The next morning The female mouse’s vaginal opening was inspected. Those who saw the semen plug with the naked eye indicated conception and began to calculate the gestational age (day 0). If the test is negative, continue to cage until the test is positive.

[0061] (2) The pregnant mice with a gestational age of 15-19 days are anesthetized with 10% chloral hydrate. After anesthesia, they are immediately put in 75% sterile alcohol and soaked for 15 minutes, and then moved into the cells. The pregnant mice are placed with the abdomen facing up and the limbs fixed. .

[0062] (3) Use sterile ophthalmic scissors...

Embodiment 2

[0099] Example 2 Comparison of extraction, separation and culture of endothelial progenitor cells from lung, heart and liver of fetal mice

[0100] The method of extraction, separation and culture is as follows:

[0101] (1) Take the pregnant SD rat, soak it with 75% ethanol immediately after anesthesia, move it into the cell, and fix the rat's limbs on the foam board on the work surface with a pin.

[0102] (2) Open the abdomen, carefully take out the uterus and fetus, move them into the ultra-clean table, remove the placenta, amniotic fluid, amniotic membrane, etc. from the fetus, and place the fetus in a petri dish with pre-cooled PBS containing double antibodies.

[0103] (3) Wash twice with PBS. Carefully remove the heart, lungs, liver and other organs of the fetus.

[0104] (4) The various organs are divided into small pieces, and each tissue fragment is digested with 0.25% pancreatin, and a stepwise digestion method is used until the tissue is complete.

[0105] (5) After the dig...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for separating and culturing lung-derived endothelial progenitor cells of fetal rats. The method comprises the steps of putting the pregnant uterus of a pregnant rat into a pre-cooled phosphate buffer solution (PBS) with both penicillin resistance and streptomycin resistance for soaking, taking out the fetal rat and putting into a plate coated with the pre-cooled PBS with both penicillin resistance and streptomycin resistance, and washing; taking out lung tissues, putting into the PBS and crushing, digesting the obtained crushed tissues by using 0.25% tyrisin,culturing the digested tissues by using a Dulbecco's modified eagle medium (DMEM) containing 10% fetal bovine serum (FBS), carrying out centrifugal separation, taking precipitate obtained by means ofthe centrifugal separation, suspending by using an EBM-2 culture medium which contains growth factors and 2% FBS, inoculating suspended matter into a six-well plate in a cell concentration of 10<6> / ML, and putting the six-well plate into a constant temperature incubator for culturing; when the primary cells grow and fuse to 80-90%, digesting with 0.25% tyrisin for subculturing, and completing theculture until the cells show a uniform morphology.

Description

Technical field [0001] The present invention belongs to the technical field of cell culture, and specifically relates to a method for extracting, separating and culturing endothelial progenitor cells, in particular to a method for extracting, separating and culturing endothelial progenitor cells from fetal mice. Background technique [0002] Endothelial progenitor cells (ENDOTHELIAL PROGENITOR CELLS, "EPCS" for short) are vascular endothelial precursor cells that can differentiate into mature endothelial cells, also known as hemangioblasts. When the body is ischemic, EPCS can mobilize from the bone marrow into the peripheral blood, reach the ischemic tissue and participate in angiogenesis. Injecting EPCS cultured and amplified in vitro into the body can increase the quantity and quality of EPCS in the circulation and promote angiogenesis in the ischemic area. Therefore, EPCS transplantation has become a new idea for the treatment of ischemic vascular disease. [0003] For the ext...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0692C12N2509/00
Inventor 计晓娟赵晓东何灿粲李亚男朱旭杨海燕龚婷李丽玲曹丽娜
Owner CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV