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Culture method for heterocephalus glaber cardiac muscle cell

A technology of cardiomyocytes and culture method, which is applied in the field of nude mole rat cardiomyocyte cultivation, can solve the problems that naked mole rat cardiomyocytes have not been reported, and achieve the effects of simple purification steps, good stability, and easy operation

Active Publication Date: 2016-04-20
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Abstract
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  • Application Information

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Problems solved by technology

[0004] However, there is no report on a culture method of naked mole rat cardiomyocytes

Method used

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  • Culture method for heterocephalus glaber cardiac muscle cell
  • Culture method for heterocephalus glaber cardiac muscle cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] (1) Add 6ml of 0.08% trypsin to 10 newborn 1-2 day old naked mole rat myocardial tissue blocks, place at 35°C, 5% CO 2 After digesting in the cell culture incubator for 6 minutes, discard the supernatant, repeat the above operation and digest again;

[0047] (2) Add 6ml of mixed digestive solution (final concentration of trypsin is 0.08%, final concentration of type II collagenase is 0.1%) to the cell pellet obtained from the above operation, gently pipette the suspension of myocardial tissue, and place it at 35°C in O 2 The volume concentration is 5%, CO 2 Digest for 6 minutes in a cell culture incubator with a volume concentration of 5%, and let stand;

[0048] (3) Collect the supernatant in step (2), filter it with a 200-mesh cell sieve, immediately add an equal amount of stop solution (DMEM low-glucose medium containing 10% FBS) to stop the digestion, and place it in a refrigerated centrifuge at 4°C , 1000rpm, and centrifuge for 5 minutes, the resulting cell pelle...

Embodiment 2

[0053] (1) Add 6.5 ml of 0.08% trypsin to 12 newborn naked mole rat myocardial tissue blocks at the age of 1-3 days, place at 35°C, O 2 The volume concentration is 5%, CO 2 After digesting for 7 minutes in a cell culture incubator with a volume concentration of 5%, discard the supernatant, repeat the above operation and digest again;

[0054] (2) Add 6.5ml of mixed digestive solution (final concentration of trypsin is 0.08%, final concentration of type II collagenase is 0.1%) to the cell pellet obtained from the above operation, gently pipette the myocardial tissue suspension, and place at 35°C, o 2 The volume concentration is 5%, CO 2 Digest for 7 minutes in a cell culture incubator with a volume concentration of 5%, and let stand;

[0055] (3) Collect the supernatant in step (2), filter it with a 200-mesh cell sieve, immediately add an equal amount of stop solution (DMEM low-glucose medium containing 10% FBS) to stop the digestion, and place it in a refrigerated centrifug...

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Abstract

The invention belongs to the technical field of cell culture, and particularly relates to a culture method for a heterocephalus glaber cardiac muscle cell. In the culture method, the operation steps of separating and purifying the heterocephalus glaber cardiac muscle cell are simple, cell attachment is fast, the survival rate is high, and a cell culture system is high in stability. The culture method is an ideal culture method for the heterocephalus glaber primary cardiac muscle cell, can basically meet the requirements for heterocephalus glaber cardiac muscle cell related experiment research in all research fields such as medical science, biology, pharmacy and bioscience and is suitable for application and popularization in general laboratories.

Description

technical field [0001] The invention belongs to the technical field of cell culture, in particular to a method for culturing naked mole rat cardiomyocytes. Background technique [0002] Naked mole rat (Heterocephalus glaber, Nakedmole-rat) is a new type of experimental animal with great research value. It has many unique biological characteristics such as hypoxia resistance, anti-aging and anti-tumor. It plays an important role in medical and life science research fields such as science and has an irreplaceable position. Especially in the study of hypoxia resistance, it provides an ideal research carrier and scientific research tool for the study of hypoxia-related cardiovascular diseases. Cardiomyocytes are the basic unit of the heart and the main source of cells for research on physiology, pharmacology, pathology, toxicology and energy metabolism. They are also one of the main sources of seed cells for myocardial tissue engineering. At present, due to the precious and ra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0657C12N2509/00C12N2525/00
Inventor 崔淑芳余琛琳汤球林丽芳赵善民丛薇蔡丽萍徐晨
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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