Primary separation culture method for bone marrow mesenchymal stem cells of naked mole rats

A bone marrow mesenchymal, isolation and culture technology, which is applied in the field of primary isolation and culture of naked mole rat bone marrow mesenchymal stem cells, can solve the problems that are not conducive to obtaining higher activity and quantity of naked mole rat bone marrow mesenchymal stem cells, and are not suitable for use. Animals with too many red blood cells, limited scope of promotion, etc., to achieve the effect of strong cell proliferation, low loss, and improved purity

Active Publication Date: 2017-08-11
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Claims
  • Application Information

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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 a...

Method used

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  • Primary separation culture method for bone marrow mesenchymal stem cells of naked mole rats
  • Primary separation culture method for bone marrow mesenchymal stem cells of naked mole rats
  • Primary separation culture method for bone marrow mesenchymal stem cells of naked mole rats

Examples

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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, and particularly discloses a primary separation culture method for bone marrow mesenchymal stem cells of naked mole rats. The primary separation culture method includes steps of 1), washing bone marrow of the naked mole rats; 2), sieving bone marrow washed substances by the aid of cell strainers; 3), breaking red blood cells; 4), carrying out direct plate laying. The primary separation culture method for the bone marrow mesenchymal stem cells of the naked mole rats has the advantages that the primary separation culture method is established for the bone marrow mesenchymal stem cells of the naked mole rats, biological characteristics of the bone marrow mesenchymal stem cells cultured by the aid of the primary separation culture method are evaluated, the fact that the bone marrow mesenchymal stem cells with extremely high purity and differentiation capacity can be obtained by the aid of the primary separation culture method is proved, and accordingly intensive study on characteristics of stem cells of the naked mole rats and exploration on characteristics of somatic cells, which are difficult to separate but can be obtained from the stem cells by means of transformation, of the naked mole rats can be possibly carried out.

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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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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