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Method for extracting and culturing Sprague Dawley (SD) rat marrow stroma cell used in autologous experiment

A technology of bone marrow stromal cells and culture methods, which is applied in the field of extraction and culture of rat bone marrow stromal cells, and can solve the problem that there is no autologous transplantation model for rat bone marrow stromal cells

Inactive Publication Date: 2010-03-17
BEIJING TIANTAN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Technical problem to be solved: In order to solve the problem that there is no suitable autologous transplantation model in the study of rat bone marrow stromal cells, the present invention is a different method for extracting and culturing rat bone marrow stromal cells

Method used

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  • Method for extracting and culturing Sprague Dawley (SD) rat marrow stroma cell used in autologous experiment
  • Method for extracting and culturing Sprague Dawley (SD) rat marrow stroma cell used in autologous experiment
  • Method for extracting and culturing Sprague Dawley (SD) rat marrow stroma cell used in autologous experiment

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Embodiment Construction

[0012] 1. Extraction

[0013] SD rats (rats aged 2-4 months) were anesthetized with 10% chloral hydrate intraperitoneally, the tail was partially disinfected, the tail was cut off along the sacrum and the tail with tissue scissors, bipolar electrocoagulation was used to stop bleeding, and local bandage was applied to rats. After waking up from anesthesia, they were put back into the breeding room and reared individually. After the clipped tail was put into 75% alcohol and soaked for 10 minutes, it was placed in another beaker and soaked in 75% alcohol for another 5 minutes. After taking it out, dissect and separate 1-6 caudal vertebrae under aseptic conditions, put them into another small beaker of 75% alcohol and soak for 3 minutes, and rinse the alcohol with D-Hank's solution. Take the first caudal vertebra under aseptic conditions, cut open the two ends of the bone marrow cavity, insert a 2.5ml syringe into the bone marrow cavity, wash once with D-Hank's solution with 1% h...

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Abstract

The invention discloses a method for extracting and culturing rat marrow stroma cells, relating to the technical field of biotic experiments. The invention is characterized in that the rat marrow stroma cells are extracted from the caudal vertebra of a rat. The rinsing step of the method comprises the following processes: rinsing by using a D-Hank's solution added with 1 percent heparin; dripping0.25 percent trypsogen+EDTA solution; neutralizing by a 15-20 percent fetal calf serum alpha-MEM culture medium; and repeatedly rinsing a marrow cavity by using the D-Hank's solution. After the marrowstroma cells are extracted by the method, the rat grows well and each physiological function is not influenced, therefore, the invention provides reliable experimental rats for carrying out various autologous transplant experiments by using marrow stroma cells.

Description

technical field [0001] The invention relates to the technical field of biological experiments. Specifically, the present invention relates to a method for extracting and culturing rat bone marrow stromal cells. After the bone marrow stromal cells of the rats are extracted by the method, the rats grow well, and various physiological functions are not affected. Provide reliable laboratory rats for various autologous transplantation experiments with bone marrow stromal cells. Background technique [0002] Bone marrow stromal cells are tissue stem cells that exist in the bone marrow stroma. They have the potential to differentiate into at least seven types of tissue cells, such as osteoblasts, chondrocytes, adipocytes, and myoblasts. Multipotent bone marrow stromal cells provide a source for the self-repair of bone, cartilage, muscle and other tissues (1、2、3) . In recent years, it has been found that it can also differentiate into astrocytes, oligodendrocytes and neurons, an...

Claims

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Application Information

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IPC IPC(8): C12N5/06
Inventor 李博张亚卓
Owner BEIJING TIANTAN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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