Immortalized Human Skeletal Muscle Myoblasts

Overview

Immortalized Human Skeletal Muscle Myoblasts is a immortalized cell line supplied in frozen format and associated with Human skeletal muscle biology.

Key elements and design rationale

• Model identity: Immortalized Human Skeletal Muscle Myoblasts is supplied as an immortalized cell line derived from Human skeletal muscle.
• Growth properties: Adherent, multipolar
• Growth conditions: For optimal cell culture, we recommend using PriCoat™ T25 Flasks (G299) or coating your preferred vessels with Applied Cell Extracellular Matrix (G422). PriGrow III (TM003) + 10% FBS (*Regular) + 10 ng/ml human EGF (Z100139) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. *Do not heat-inactivate Note: Maintain at high density (minimum 60% confluency) during first few passages for optimal recovery. Proliferation is expected to be slow initially; however, after a recovery period of 1 to 2 weeks, the cells typically resume a more robust growth rate, with a doubling time of approximately 24 to 36 hours.
• Product format: Frozen, BSL-2

This cell-based model is generally used in skeletal muscle biology, phenotype comparison, and assay development studies. Donor/background information is available for contextual interpretation.

Biological background

This model supports studies in skeletal muscle biology, phenotype comparison, and assay development. It can be used to examine morphology, growth behavior, and experimental responses in cultured cells. Donor/background information provided for this product: Male, 7, Caucasian.

Research relevance and current trends

• Cultured cell-line models remain central to in vitro studies of phenotype, signaling, and pathway regulation under controlled conditions.
• Researchers commonly compare morphology, growth rate, and marker expression across media formulations, treatments, or time courses.
• Interpretation is generally strengthened by using matched controls, consistent passage handling, and appropriate culture surfaces.

Common research applications

• Routine expansion and maintenance of a defined cell model for downstream in vitro experiments.
• Phenotype, signaling, or marker-expression studies performed under standardized culture conditions.
• Cell-based assay development in which passage number, growth surface, and medium composition are tracked as experimental variables.

Changes in morphology, growth rate, viability, or reporter signal are typically interpreted together with passage history, culture matrix, and the specified growth conditions for the model.

Notes for experimental interpretation

• Morphology, doubling behavior, and reporter or marker output can shift with passage number, substrate choice, and medium composition; these variables should be recorded alongside experimental readouts.
• Matched controls such as parental cells, untreated cultures, or parallel cultures maintained under identical conditions help distinguish background effects from biology of interest.

Culture and product details

• Growth Conditions: For optimal cell culture, we recommend using PriCoat™ T25 Flasks (G299) or coating your preferred vessels with Applied Cell Extracellular Matrix (G422). PriGrow III (TM003) + 10% FBS (*Regular) + 10 ng/ml human EGF (Z100139) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. *Do not heat-inactivate Note: Maintain at high density (minimum 60% confluency) during first few passages for optimal recovery. Proliferation is expected to be slow initially; however, after a recovery period of 1 to 2 weeks, the cells typically resume a more robust growth rate, with a doubling time of approximately 24 to 36 hours.
• Seeding Density (cells/cm²): 20,000 - 30,000
• 3D Culture Conditions: Procedure Preparation (before 3D Culture): Thaw cells following "Thawing Protocol" and "Growth Conditions". Expand cells for one passage, then seed for spheroid generation (3D culture). Subculture Protocol:Aspirate the supernatant and wash cells with 1X PBS (pH 7.4) (G5000). Dissociate cells using 1–1.5 ml of Gentle Dissociation Solution (TM080). Observe the cells under a microscope to confirm detachment (typically within 2–10 minutes). Cells that are difficult to detach can be put in 37 °C for several minutes to facilitate detachment.Neutralize using an equal volume of complete growth media (must contain serum) or using Trypsin Neutralizing Solution (TM069).Transfer culture suspension into sterile conical centrifuge tube (G5500), and centrifuge at 125 x g for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.Aspirate supernatant and re-suspend the pellet with pre-warmed fresh complete growth media for 3D Culture. Spheroid Generation Protocol:Count cells and viability using cell counter or hemocytometer with Trypan Blue Stain (TM071). Proceed if cells are >90% viability.Seed 500 to 10,000 cells per well in SpheroWell™ 96 Well Plate (G7540), final volume maintained at 100 μl per well. Avoid cell clumps (clumps will not generate uniform spheroids). Count as Day 0. Tip: Fill the outermost wells of the plate with 1X PBS (pH 7.4) (G5000) to prevent evaporation of media during incubation.(Recommended) Centrifuge the sealed plate at 300 x g for 5 minutes.Place in the incubator set at 37 °C, 5% CO₂. Change Media:Every 2–3 days, add 100 μl of fresh media slowly along the wall of the well to avoid disrupting spheroids at the bottom.Take out 100 μl of media slowly and discard from the well.(Recommended, but optional) Centrifuge the sealed plate at 300 x g for 5 minutes. Monitor the Cells for 3D/Spheroid Formation:Spheroid formation will appear between day 5 and 10.

Cell line sourcing and selection (species, tissue, and disease model matching) · Stable cell line engineering (overexpression, knockdown, knockout via CRISPR/Cas9, shRNA, sgRNA) · Reporter gene integration (GFP, RFP, luciferase, fluorescent/bioluminescent constructs) · Genome editing and knockin (point mutations, tagged endogenous proteins, conditional alleles) · Inducible expression systems (Tet-On/Off and regulatable constructs) · Drug resistance marker selection (puromycin, G418, hygromycin, and others) · Custom growth and media optimisation for specific assay requirements · Scale-up production for high-throughput screening campaigns · Authentication and QC services (STR profiling, mycoplasma testing, viability assessment). Talk to a Scientist or contact support@biohippo.com.