Cardiomyocytes are one of the cell groups that compose the heart. Cardiomyocytes are known as beating involuntary striated muscle cells. The "beat" which is the feature of the cardiomyocytes can be widely used for pharmacologic and electrophysiology assays, because it responds to a variety of stimuli including hormones, drugs and electricity.
Cardiomyocyte Culture Kit contains cryopreserved cardiomyocytes, culture medium and fibronectin. Cells are isolated from mouse embryos’ heart, and cardiomyocytes are enriched by removal of non-cardiomyocytes. The involuntary beat of the cells can be seen in culture using Cardiomyocyte Culture Kit (PMC-CMC12-COS).
|Product Name / Code No.||Size||Quantity||Storage Conditions||Shelf Life|
|2 x 106 cells/vial
|1||Liquid Nitrogen||1 year|
|125 mL||1||-20°C Freezer||6 months|
< 1-1. Cultured with the 24well-plate >
A seeding density of 2.0×105 cells/well is recommended.
1. Thaw the Culture medium in at 37°C water bath with gentle shaking.
2. Quickly thaw the Cardiomyocyte vial for 1min 15sec in a 37°C water bath.
3. Transfer the vial contents of thawed cells into a 15 mL centrifuge tube containing 4.5 mL of Culture Medium.
4. Gently mix gently the cell suspension by slow pipetting up and down, and adjust cell density to 2.0×106 cells/5 mL solution in the tube.
5. Transfer 0.5 mL of cell suspension to each well of fibronectin-coated 24-well plate.
6. Incubate the plate at 37oC in a 5 % CO2 humidified incubator.
7. The next day, gently add 0.5 mL of fresh pre-warmed culture medium to each well.
8. Exchange the medium with fresh and pre-warmed culture medium every day or every other day until the culture reaches 80-100% of cell confluent.
* Culture reaches its confluent within 3-5 days, and cardiomyocytes begin beating.
* Do NOT use cold culture medium. Please pre-warm culture medium at 37oC before use to ensure the viability of cardiomyocytes.
Fig. 1 Morphology of cardiomyocyte
< 1-2. Culture adding 5-Bromo-2’-deoxyuridine >
It is recommended to supplement 5-Bromo-2’-deoxyuridine (BrdU) to culture in order to increase the purity of cardiomyocytes (BrdU prevents non-cardiomyocytes from developing). Preparation of BrdU: Dissolve BrdU (Sigma Cat. No. B-9285 or the similar product) in culture medium to a concentration of 10mM and filter sterilization. Fresh BrdU solution should be used for cardiomyocyte culture.
Prepare cell suspension as described above in steps 1-4 (<1-1. Cultured with the 24well-plate>).
Transfer 0.5 mL of cell suspension to each well of fibronectin-coated 24-well plate and add 5 μl of 10mM BrdU to each well (0.1 mM BrdU-final concentration).
Incubate the plate at 37oC in a 5 % CO2 humidified incubator.
The next day, exchange the medium with fresh 0.5 mL of the medium containing 0.1 mM BrdU.
On day 3 of the culture, replace the medium with fresh 0.5 mL of the medium without BrdU.
< Prepare a fibronectin coated Culture 24-well plate >
Thaw fibronectin solution.
Add 0.5 mL of fibronectin solution to each well. (Add 0.25 mL/cm2 of solution to each well in other types of culture plates.)
Leave the plate at 37oC in 5% CO2 humidified incubation overnight.
Aspirate remaining material and rinse plates with sterile water twice. Plates are ready for use.
Cardiomyocyte Culture Medium
Fibronectin coat solution
To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.