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A primary isolation and culture method of naked mole rat bone marrow mesenchymal stem cells

A bone marrow mesenchymal, isolation and culture technology, applied in the field of primary isolation and culture of naked mole rat bone marrow mesenchymal stem cells, can solve the problem of obtaining high activity and quantity of naked mole rat bone marrow mesenchymal stem cells, inapplicable Animals with too many red blood cells, limited promotion range, etc., achieve the effect of strong cell proliferation ability, low loss, and improved purity

Active Publication Date: 2021-05-07
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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AI Technical Summary

Problems solved by technology

The cost of flow cytometry and immunomagnetic bead separation is high, and the scope of promotion is limited; the cell viability obtained by density gradient centrifugation is low when isolating bone marrow mesenchymal stem cells of small animals; the cell viability obtained by direct attachment method is high , but not suitable for animals with excess erythrocytosis due to erythrocytes affecting the attachment of target cells
Naked mole rats are close to mice in size, adapt to low oxygen environment, and have high red blood cell content. The current culture method is not conducive to obtaining high activity and quantity of naked mole rat bone marrow mesenchymal stem cells

Method used

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  • A primary isolation and culture method of naked mole rat bone marrow mesenchymal stem cells
  • A primary isolation and culture method of naked mole rat bone marrow mesenchymal stem cells
  • A primary isolation and culture method of naked mole rat bone marrow mesenchymal stem cells

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Embodiment 1

[0037] Embodiment 1 Method for Isolation and Culture of Naked Mole Rat Bone Marrow Mesenchymal Stem Cells

[0038] (1) Take an adult naked mole rat, euthanize it by cervical dislocation, soak it in 75% ethanol, and move it into a clean bench. The two hind limbs were taken under aseptic conditions, the skin and muscles were peeled off, the femur and tibia were separated, and the cartilage tissues at both ends were removed. Gently insert a 1mL syringe into the bone marrow cavity, flush the bone marrow cavity with washing solution, and obtain bone marrow flushing fluid.

[0039] (2) Pass the bone marrow fluid through a 200-mesh cell sieve to make a single-cell suspension;

[0040] (3) Centrifuge the above cell suspension at 100 g for 5 min, discard the supernatant, and collect the cell pellet;

[0041] (4) Resuspend the cell pellet in 2 mL of erythrocyte lysate, lyse the erythrocytes for 1-2 minutes, add an equal volume of stop solution to terminate the lysis, and discard the s...

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Abstract

The invention relates to the field of stem cell culture, in particular to a method for primary isolation and culture of naked mole rat bone marrow mesenchymal stem cells, comprising the following steps: 1) washing the naked mole rat bone marrow; 2) passing the bone marrow flushed out through a cell sieve; 3) Destroy red blood cells; 4) direct plating. The present invention establishes a method for isolating and culturing naked mole rat bone marrow mesenchymal stem cells, and evaluates the biological characteristics of the cells cultured by the method, confirming that the culture method can obtain naked mole rat bone marrow with high purity and differentiation ability Mesenchymal stem cells provide the possibility to further study the characteristics of naked mole rat stem cells and explore the characteristics of naked mole rat somatic cells that are not easy to separate but can be transformed by stem cells.

Description

technical field [0001] The invention relates to the field of stem cell culture, in particular to a method for primary isolation and culture of naked mole rat bone marrow mesenchymal stem cells. Background technique [0002] The naked mole rat is a digging rodent distributed in parts of East Africa, and belongs to the class Mammalia and Rodentia in taxonomy. Naked mole rats live their entire lives in the dark, underground, adapt to low-oxygen environments, and have the longest lifespan among rodents; they are super immune to cancer. Longevity, anti-aging, hypoxia resistance, anti-tumor and other properties of naked mole rats have attracted widespread attention from scientists, and research results on these properties have been published in famous journals such as Nature and PNAS. [0003] Bone marrow mesenchymal stem cells (BMSCs) are adult stem cells isolated from bone marrow with high self-renewal ability and multilineage differentiation potential. Although mesenchymal st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0663C12N2501/73
Inventor 崔淑芳丛薇李周桐杨文静余琛琳赵善民林丽芳程继帅刘攀徐晨
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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