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Method for obtaining mesenchymal stem cells from mouse bone marrow

A bone marrow mesenchymal and mesenchymal stem cell technology, applied in the field of mesenchymal stem cell preparation, can solve the problems of low efficiency of mesenchymal stem cells, and achieve the effects of high efficiency, convenient research and testing, and good in vitro differentiation potential

Pending Publication Date: 2020-02-11
深圳市蓝思人工智能医学研究院 +1
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  • Abstract
  • Description
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Problems solved by technology

The efficiency of this method to obtain mesenchymal stem cells is low, and a certain proportion of CD45-positive macrophages will be mixed in the cultured cells

Method used

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  • Method for obtaining mesenchymal stem cells from mouse bone marrow
  • Method for obtaining mesenchymal stem cells from mouse bone marrow
  • Method for obtaining mesenchymal stem cells from mouse bone marrow

Examples

Experimental program
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Effect test

Embodiment

[0035] 1. Isolation of mouse bone marrow mesenchymal stem cells by enzymatic hydrolysis

[0036]In this example, commercially purchased C57 female or male mice of 6 to 8 weeks were sacrificed with carbon dioxide, and the receding femurs and tibias on both sides of the mice were obtained by dissecting them with mouse dissecting instruments. After the excess muscle is removed, the bone marrow of the femur and tibia is blown out of the bone marrow cavity with a dispensing needle to keep the integrity of the bone marrow from being damaged. Then, the bone marrow of each mouse was digested twice with 700 μL of HBSS buffer containing 3 mg / mL type 1 collagenase and 20 U / mL dispase (hereinafter referred to as digestion solution) at 37 ° C on a constant temperature shaker at 220 rpm / min. Specifically, after one digestion, settle at room temperature for 5 minutes, transfer the supernatant to 10 mL of PBS for neutralization and digestion, and then add 700 μL of digestion solution for the ...

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Abstract

The present invention discloses a method for obtaining mesenchymal stem cells from mouse bone marrow. The method for obtaining mesenchymal stem cells from mouse bone marrow comprises steps of obtaining intact non-destructive bone marrow from a femur or a tibia of a mouse, and then digesting the obtained intact bone marrow with collagenase to obtain mesenchymal stem cells. The method for preparingmesenchymal stem cell from mouse bone marrow improves efficiency of preparing mesenchymal stem cells from mouse bone marrow and the prepared mesenchymal stem cells have better differentiation potential in vitro.

Description

technical field [0001] The present application relates to the field of preparation methods of mesenchymal stem cells, in particular to a method for obtaining mesenchymal stem cells from mouse bone marrow. Background technique [0002] Mesenchymal stem cells are a highly heterogeneous group of stromal stem cells. A variety of adult tissues and organs contain mesenchymal stem cells. Mesenchymal stem cells were first isolated from bone marrow, and can form spindle-shaped mononuclear cells with clone-forming ability when cultured in vitro. When cultured in vitro, mesenchymal stem cells can not only serve as supporting cells for hematopoietic stem cells, but also have a variety of differentiation potentials, and can differentiate into osteoblasts, adipocytes and chondrocytes. In addition to in vitro functions, a large number of studies have shown that mesenchymal cells are also an important part of the microenvironment of hematopoietic stem cells in vivo, and also play an impor...

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 赵萌娄琪牟丽莎陆赢蒲祖辉
Owner 深圳市蓝思人工智能医学研究院
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