H9C2 rat cardiomyocytes

H9C2 rat cardiomyocytes

Cell identification

Species identification has been passed

Cell source

National Resource Library

Cell background

B. Kimes and B. Brandt subcloned H9c2 (2-1) cell line from a cloned cell line derived from BD1X rat embryonic heart tissue, which exhibits many skeletal muscle characteristics. The myoblasts in this cell line can fuse to form multinucleated myotubes and respond to acetylcholine stimulation. If the serum concentration in the culture medium drops to 1%, fusion occurs quickly.

Cell characteristics

1) Source: Heart

2) Morphology: Myoblasts, adherent growth

3) Content:>1x106 cell count

4) Specification: Packaging in T25 bottles or 1mL cryovials

5) Purpose: For scientific research purposes only.

Cultivation conditions:

1) Prepare DMEM (containing 1.5g/L NaHCO3, recommended: awcell-128-0001) or (GIBCO, item number 12800017, with added NaHCO3 1.5g/L) culture medium; High quality fetal bovine serum, 10%; Double antibody 1%.

2) Cultivation conditions: Gas phase: air, 95%; Carbon dioxide, 5%. Temperature: 37 degrees Celsius, humidity in the incubator is 70% -80%.

L Precautions:

Attention: Rat cardiomyocytes H9C2 grow well in DMEM (containing 1.5g/L NaHCO3) medium. Most brands of DMEM contain high concentrations of NaHCO3 (3.7g/L). If DMEM (3.7g/L NaHCO3) medium is used to culture cells, the CO2 concentration needs to be increased (7% -10%).

For the passage of adherent cells, the following methods can be referred to:

1. Discard the culture supernatant and rinse the cells 1-2 times with PBS that does not contain calcium or magnesium ions.

2. Add 0.25% (w/v) trypsin 0.53 mM EDTA to culture bottles (1-2mL for T25 bottles and 2-3mL for T75 bottles), digest in a 37 ℃ incubator for 1-2 minutes (digestion time can be appropriately extended for difficult to digest cells), and then observe the digestion of cells under a microscope. If most of the cells become round and fall off, quickly take them back to the operating table, tap the culture bottle a few times, and add 3-4ml of culture medium containing 10 μ S to terminate digestion.

3. Gently mix and aspirate, centrifuge at 1000RPM for 3-5 minutes, discard the supernatant, add 1-2mL of culture medium and blow evenly. Divide the cell suspension into new T25 bottles in a ratio of 1:2, add 6-8ml of new * culture medium prepared according to the instructions to maintain the growth vitality of the cells, and subsequently subculture according to the actual situation in a ratio of 1:2~1:5.

3) Cell cryopreservation: It is recommended to freeze a batch of cell seeds during the first 3 generations of culture after receiving the cells for subsequent experimental use.

Transportation mode:

Low temperature: (1) Transport in 1mL cryovials packaged with dry ice. After receiving, store in a -80 degree freezer overnight and transfer to liquid nitrogen or directly resuscitate. If you find that the dry ice has evaporated completely, the cryovial cap has fallen off, is damaged, or the cells are contaminated, please contact us immediately.

At room temperature (2), T25 bottles of revived surviving cells will be shipped at room temperature and processed according to the procedures for cell reception upon receipt.

Biosafety:

1. All animal cells are considered to have potential biological hazards and must be operated in a secondary biosafety platform. Please pay attention to protection, and all waste liquids and containers that have come into contact with these cells must be sterilized before disposal.

2. It is recommended to always use protective gloves, clothing, and a face mask when reviving frozen cells. Attention: The cryotube immersed in liquid nitrogen will leak and gradually fill with liquid nitrogen. When thawing, the conversion of liquid nitrogen into gas phase may cause the container to explode or the lid to be blown off with dangerous force, resulting in flying debris and causing personal injury.