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Method for extracting mesenchymal stem cells and application

A bone marrow mesenchymal and stem cell technology, applied in the field of cell engineering, can solve the problems of increasing the probability of pollution, cell loss, affecting stem cell adhesion, and loss of culture medium nutrition, so as to reduce the risk of cell pollution, low cost, and stem cell low loss effect

Inactive Publication Date: 2018-09-14
THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method is cumbersome, and the filtration and centrifugation steps increase the chance of contamination and the loss of cells
After centrifugation, a large number of red blood cells and stem cells are mixed together, which affects the attachment of stem cells, resulting in a low proportion of stem cells in primary cells and loss of nutrients in the medium

Method used

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  • Method for extracting mesenchymal stem cells and application
  • Method for extracting mesenchymal stem cells and application
  • Method for extracting mesenchymal stem cells and application

Examples

Experimental program
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Embodiment 1

[0046]SD rats, about 100g, male or female, were anesthetized by intraperitoneal injection of 10% chloral hydrate 3-4ml / kg, killed by cervical dislocation, and their fur was disinfected with povidone iodine. The femur and tibia were separated in a sterile ultra-clean bench, washed once with sterile PBS, and then soaked in DMEM medium (Hyclone) containing 10% fetal bovine serum. The metaphysis of the femur and tibia were subtracted using sterile scissors, exposing the medullary cavity. Use a 1ml syringe with a 4.5 needle to draw DMEM medium (Hyclone) containing 10% fetal bovine serum, gently blow the contents of the bone marrow cavity into a well of a six-well plate (Corning), try to keep the contents intact, and use the medium repeatedly Beat the bone marrow cavity until the bone marrow cavity turns white. At this time, the contents of the bone marrow cavity can be seen as lumps. The washing solution was sucked and centrifuged at 1200 rpm for 5 minutes, the supernatant was di...

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Abstract

The invention relates to a method for extracting mesenchymal stem cells. The method includes the steps: performing pretreatment to obtain a thigh bone and a shin bone; flushing the thigh bone and theshin bone, extracting marrow cavity contents to obtain flushing liquid containing crumby marrow cavity contents; and collecting stem cells in the flushing liquid; blowing and beating remaining crumbymarrow cavity contents to obtain single-cell suspensions, and collecting stem cells in the single-cell suspensions; grafting and culturing the acquired stem cells. The method can be used for extracting mesenchymal stem cells of rats, and the acquired mesenchymal stem cells are applied to research of cell biology. The method has the advantages of simplicity in operation, low cost, pollution probability and stem cell loss, more cell extraction amount, high purity, good repeatability and the like, and the method is of great significance for biological research in the field.

Description

technical field [0001] The present invention relates to the technical field of cell engineering, and more specifically, relates to a method for extracting bone marrow mesenchymal stem cells and the application of the method in extracting rat bone marrow mesenchymal stem cells. Background technique [0002] Rat bone marrow mesenchymal stem cells are one of the main seed cells for stem cell research. It is capable of osteogenic, adipogenic and chondrogenic differentiation, and has a strong proliferative capacity. The sources of rat bone marrow mesenchymal stem cells used in the study were mainly purchased commercial cell lines or extracted from rat femur and tibia. [0003] At present, the main method for extracting rat bone marrow mesenchymal stem cells is the whole bone marrow adherence method. This method mainly utilizes the property of bone marrow mesenchymal stem cells to adhere easily to separate them from other blood cells. The main step is to aspirate the bone marro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0663C12N2509/00
Inventor 蓝蔚仁孙超左睿李海音蒋长青潘赛李长青
Owner THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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