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Method of high-effectively obtaining mesenchymal stem cells from compact bone of mice

A mesenchymal stem cell and mouse technology, applied in the field of mesenchymal stem cell acquisition, can solve the problems of decreased cell number and differentiation ability, difficulty in obtaining a large number of mouse bone marrow cells, and contamination of non-MSCs adherent cells, so as to shorten the time spent, Fast and efficient acquisition, the effect of increasing cell volume

Active Publication Date: 2017-10-24
ZHEJIANG UNIV
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Problems solved by technology

[0004] Current research on stem cells mostly focuses on bone marrow-derived MSCs, but because it is very difficult to obtain a large number of mouse bone marrow cells, they are easily contaminated by non-MSCs adherent cells during the culture process, and the number of cells and their differentiation ability increase with the age of mice. There is a clear downward trend

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  • Method of high-effectively obtaining mesenchymal stem cells from compact bone of mice
  • Method of high-effectively obtaining mesenchymal stem cells from compact bone of mice
  • Method of high-effectively obtaining mesenchymal stem cells from compact bone of mice

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Embodiment Construction

[0042] At first the source of the mouse humerus, tibia and femur used in the present invention is described:

[0043] The mouse humerus, tibia and femur used in the present invention are all taken from the discarded natural dead mice in the laboratory: choose the BALB / c young mouse corpse or the C57BL / 6 young mouse corpse with a body weight of 6-8g at the age of 2-3 weeks , after cleaning with 100ml of 70% ethanol, take out the humerus, tibia and femur in the biological safety cabinet for subsequent tests. During the implementation of the present invention, there is no operation of killing surviving mice, and there is no traumatic or interventional treatment such as dissection and excision of living mice.

[0044]In the present invention, the MEM culture fluid is commercially available, and is HyClone TM Produced MEM AlphaModification (1×) medium, catalog number SH30265.01 (official website of reagent manufacturer: www.gelifescience.com / hyclone).

[0045] The technical solut...

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Abstract

The invention relates to methods of obtaining mesenchymal stem cells and is to provide a method of high-effectively obtaining mesenchymal stem cells from compact bone of mice. The method of high-effectively obtaining mesenchymal stem cells of mice includes the steps of: separating mesenchymal stem cells of mice, performing primary culture, and performing amplification culture to the mesenchymal stem cells of mice. The method greatly reduces the time of cell culture and can greatly increase quantity of produced cells, and is less in pollution of non-MSCs adherent cells. The invention, for the first time, discloses a multi-time bone sheet adhesion culture method, which can increase the yield of cells by 3-5 times. Through systematic researching on biological characteristics, such as growth status, surface markers, multidirectional differentiative potential and the like, of the MSCs from the compact bone of mice, it is proved that after 10 times of amplification culture in vitro, the cells still have strong characters of stem cells and can satisfy demands in experiment and researching. The method can quickly and high-effectively produce the stem cells. The obtained stem cells has high proliferation and growth speed and high differentiative potential.

Description

field of invention [0001] The invention relates to a method for obtaining mesenchymal stem cells, in particular to a method for efficiently obtaining mesenchymal stem cells from mouse compact bone. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of stem cells, a group of fibroblast-like cells that adhere to the wall, have multi-directional differentiation ability, and can produce various cytokines and growth factors; they have hematopoietic support, Immunomodulatory and anti-inflammatory functions have great scientific and practical value in the field of regenerative medicine. [0003] MSCs are generally considered not a single type of cell, but a mixed cell population, so the antigen expression on its surface is not specific. MSCs markers include Sca-1, CD29, CD44 and CD105 and other surface markers positive. Excludes CD31, CD34, CD45 and MHC-1A. In addition, identification of differentiation ability, including osteogenic and adipogenic ability, is...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0663C12N2509/00
Inventor 曹红翠李兰娟俞炯曾浔潘巧玲
Owner ZHEJIANG UNIV
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