H9c2(2-1)

€375.00*

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Product number: 305203

Safety data sheet

Product sheet

Description	Introducing H9c2(2-1) Cells: A Versatile Cell Model for Cardiomyocyte Research H9c2(2-1) cells are a specialized subclone derived from the original clonal cell line obtained from embryonic BD1x rat heart tissue. These cells exhibit remarkable properties similar to skeletal muscle cells, making them an ideal model for studying cardiac muscle function. When stimulated by acetylcholine, the myoblastic cells in the H9c2(2-1) cell line can fuse and form multinucleated myotubes. These cells are derived from the embryonic heart tissue of the Rattus norvegicus (Rat) species, specifically the cardiac muscle. As an alternative to primary cardiomyocytes, H9c2 cells offer a valuable model for cardiac research. By reducing the serum concentration in the cell culture media and introducing all-trans-retinoic acid (RA), the H9c2 cells can be induced to differentiate into a cardiac-like phenotype. This differentiation process results in multinucleated cells with reduced proliferative capacity, resembling cardiomyocytes. In a groundbreaking study, the transcriptional analysis of H9c2 cells was conducted in two differentiation states: embryonic cells and differentiated cardiac-like cells. The findings revealed significant upregulation of genes encoding cardiac sarcomeric proteins, calcium transporters, and associated machinery. Notably, genes associated with mitochondrial energy production, such as respiratory chain complexes subunits, mitochondrial creatine kinase, carnitine palmitoyltransferase I, and uncoupling proteins, were also observed to increase expression during differentiation. Undifferentiated H9c2 myoblasts demonstrated increased expression of genes related to pro-survival proteins and cell cycle regulation. The differentiation of H9c2 cells led to the remodelling of mitochondrial function and increased transcripts and protein levels associated with calcium handling, glycolytic and mitochondrial metabolism. The involvement of PI3K, PDK1, and p-CREB in H9c2 differentiation was also identified. Complex analysis of differently expressed transcripts further revealed significant upregulation of genes associated with cardiac muscle contraction, dilated cardiomyopathy, and other pathways specific to cardiac tissue. The H9c2 (2–1) cell line offers several advantages over primary cardiomyocytes. Primary cardiomyocytes are delicate and challenging to maintain in culture for extended periods. Moreover, their isolation requires the sacrifice of laboratory animals, raising ethical concerns. In contrast, H9c2 cells provide a reliable in vitro model for studying cardiac and skeletal muscle, enabling cell biology, electrophysiology, and toxicology research without the need for animal sacrifice. Initially isolated from the ventricular part of a BDIx rat heart, the H9c2 cell line undergoes selective serial passages, allowing the separation of different components based on their adhesion kinetics. Although not fully differentiated into adult cardiomyocytes, these cells exhibit cardiomyocyte-specific markers, demonstrating their potential for cardiac research. By culturing H9c2 cells in a low serum concentration media, they can differentiate from mono-nucleated myoblasts to myotubes, assuming an elongated shape and parallel positioning. The addition of all-trans retinoic acid (RA) to the media further induces the presence of cells with an adult cardiac muscle phenotype characterized by the overexpression of the alpha-1 subunit of L-type calcium channels. It is important to note that H9c2 cells do not possess contractile activity, even in their differentiated state. Nevertheless, these cells respond similarly to various stimuli as isolated neonatal cardiomyocytes, including developing hypertrophic responses. However, it is crucial to consider the differentiation between state
Organism	Rat
Tissue	Heart, myocardium
Synonyms	H9c2 (2-1), H9c2, H9C2

Age	Embryo
Morphology	Myoblast
Growth properties	Adherent

Citation	H9c2(2-1) (Cytion catalog number 305203)
Biosafety level	1

Receptors expressed	Acetylcholine, expressed
Protein expression	Myokinase, Creatine Phosphokinase, Myosin

Culture Medium	DMEM
Medium supplements	10% FBS, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium pyruvate
Passaging solution	Accutase
Subculturing	Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10 ml for T75 cell culture flasks). Add Accutase (1-2 ml per T25, 2.5 ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 8-10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300 g, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium.
Split ratio	1:2 to 1:4
Fluid renewal	2 to 3 times per week
Freeze medium	CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100)
Handling of cryopreserved cultures	The cells come deep-frozen shipped on dry ice. Please make sure that the vial is still frozen. If immediate culturing is not intended, the cryovial must be stored below -150 degree Celsius after arrival. If immediate culturing is intended, please follow the below instructions: Quickly thaw by rapid agitation in a 37 degree Celsius water bath within 40-60 seconds. The water bath should have clean water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water bath. A small ice clump should still remain and the vial should still be cold. From now on, all operations should be carried out under aseptic conditions. Transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open the vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of culture medium (room temperature). Resuspend the cells carefully. Centrifuge at 300 x g for 3 min and discard the supernatant. The centrifugation step may be omitted, but in this case the remains of the freeze medium have to be removed 24 hours later. Resuspend the cells carefully in 10 ml fresh cell culture medium and transfer them into two T25 cell culture flasks. All further steps are described in the subculture section.
Handling of proliferating cultures	One or two cell culture flasks come filled with cell culture medium. Collect the entire medium in 1 or 2 x 50 ml centrifuge tubes, respectively. Carefully add 5 ml of cell culture medium to each T25 cell culture flask. Control the cell morphology and confluency under the microscope. Incubate at 37 degree Celsius for a minimum of 24 hours. Spin down the collected medium at 300 x g for 3 minutes to collect the cells which may have detached during transit. If a cell pellet is visible, resuspend the cells in 5 ml of cell culture medium and transfer to a T25 cell culture flask. Incubate at 37 degree Celsius for a minimum of 24 hours.

Sterility	Mycoplasma contaminations are ruled out through PCR-based and luminescence-based mycoplasma assays. The absence of bacterial, fungal or yeast contamination is controlled through daily visual cell monitoring.

Contamination-free cells

To identify mycoplasma contaminations we perform PCR-based and luminescence-based mycoplasma assays. We further determine any bacterial or fungal contamination through our standardized manufacturing processes.

Custom projects

Besides genomic DNA, RNA, cell pellets, and cell lysates, we can offer large quantities of assay-ready cells, plated cells in multiple formats, and frozen or growing cells. Contact us to receive a quote.

Authenticated cells

Each manufactured batch of cell lines* is authenticated via STR analysis. Contact us if you require a publication-ready STR report for your cells (*human, hamster, mouse, rat, and dog cells).

HLA alleles

HLA characterization is available from more than 200 cell lines. HLA class I -A, B, C, and Class II HLA-DPA1, -DPB1, -DQA1, DQB1, and DRB1 alleles were obtained by next-generation sequencing methodologies (NGS) for class I and class II alleles.

H9c2(2-1)

General information

Characteristics

Identifiers / Biosafety / Citation

Expression / Mutation

Handling

Quality control / Genetic profile / HLA