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| Biosafety level | |
| Product format | Frozen |
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Overview
The parental L6 myoblast cell line (Cat. No.
Key elements and design rationale
- Model identity: Immortalized GLUT4-Overexpressing Rat Skeletal Myoblast Cells (L6) is supplied as an immortalized cell line derived from Rat skeletal muscle.
- Growth properties: Adherent, polygonal
- Growth conditions: Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow VIII (TM018) + 10% FBS(Regular*) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂ *Do not heat-inactivate
- 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
T0771) has been transfected with GLUT4 cDNA to give the Immortalized Rat Skeletal Myoblast Cell line (L6), or L6-GLUT4myc. The cells display a myoblast morphology and retain the potential to differentiate. In the plasma membrane, GLU4 is responsible for the activation of glucose transportation by insulin. The Immortalized Rat Skeletal Myoblast Cell line (L6) has been used to study glucose transport systems and is suitable for research pertaining to muscle physiology and metabolism. Donor/background information provided for this product: 2 day old rat pup.
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: Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow VIII (TM018) + 10% FBS(Regular*) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂ *Do not heat-inactivate
- Seeding Density (cells/cm²): 20,000 - 30,000
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🧊 Thawing Protocol
- Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.
- Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.
- Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.
- Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.
- Incubate the cells at the recommended conditions.
🔬 Subculture Protocol
Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent.
- Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.
- 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 Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.
- Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.
- Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired.
- Incubate the cells at the recommended conditions.
How should I handle live cells once I receive them?
Please refer to our Cell Handling and Thawing Guidelines for detailed instructions on receiving, thawing, and culturing live cells:
https://www.abmgood.com/immortalized-cells-documents.html
Following these guidelines will help ensure optimal cell viability and performance.
https://www.abmgood.com/immortalized-cells-documents.html
Following these guidelines will help ensure optimal cell viability and performance.
Why are these cells classified as biosafety level II?
We follow the CDC-NIH recommendations that all mammalian sourced products should be handled at the Biological Safety Level 2 to minimize exposure of potentially infectious products. This information can be found in 'Biosafety in Microbiological and Biomedical Laboratories' (1999). Your institution's Safety Officer or Technical Services will be able to make the call as to whether BioSafety Level I is possible with these cells at your site, if desired.
What is your warranty or return policy?
Our warranty and return policy is outlined in abm’s Terms and Conditions, including details on product quality, limitations, and claims.
Please refer to the following link for full information:
https://www.abmgood.com/terms
For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com.
Please refer to the following link for full information:
https://www.abmgood.com/terms
For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com.
How many times can cells divide?
The number of times cells can divide depends on the cell type:
Primary cells have a limited lifespan and will undergo a finite number of population doublings before entering senescence. The exact number varies by cell type and culture conditions.
Immortalized cell lines are capable of extended or indefinite proliferation under proper culture conditions, although growth characteristics may vary between lines.
Primary cells have a limited lifespan and will undergo a finite number of population doublings before entering senescence. The exact number varies by cell type and culture conditions.
Immortalized cell lines are capable of extended or indefinite proliferation under proper culture conditions, although growth characteristics may vary between lines.
Do I need Applied Cell Extracellular Matrix (G422) if I am using PriCoat™ flasks?
Please refer to the Growth Conditions section of your specific product page. This section will indicate whether Applied Cell Extracellular Matrix (G422), PriCoat™ flasks, or both are recommended for optimal cell attachment and growth, as requirements may vary by cell type.
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.
Kanai, F., Nishioka, Y., Hayashi, H., Kamohara, S., Todaka, M., & Ebina, Y. (1993). Direct demonstration of insulin-induced GLUT4 translocation to the surface of intact cells by insertion of a c-myc epitope into an exofacial GLUT4 domain. The Journal of biological chemistry, 268(19), 14523–14526.
Wang, Q., Khayat, Z., Kishi, K., Ebina, Y., & Klip, A. (1998). GLUT4 translocation by insulin in intact muscle cells: detection by a fast and quantitative assay. FEBS letters, 427(2), 193–197. https://doi.org/10.1016/s0014-5793(98)00423-2
Wang, Q., Somwar, R., Bilan, P. J., Liu, Z., Jin, J., Woodgett, J. R., & Klip, A. (1999). Protein kinase B/Akt participates in GLUT4 translocation by insulin in L6 myoblasts. Molecular and cellular biology, 19(6), 4008–4018. https://doi.org/10.1128/MCB.19.6.4008
Rudich, A., Konrad, D., Török, D., Ben-Romano, R., Huang, C., Niu, W., Garg, R. R., Wijesekara, N., Germinario, R. J., Bilan, P. J., & Klip, A. (2003). Indinavir uncovers different contributions of GLUT4 and GLUT1 towards glucose uptake in muscle and fat cells and tissues. Diabetologia, 46(5), 649–658. https://doi.org/10.1007/s00125-003-1080-1
Antonescu, C. N., Randhawa, V. K., & Klip, A. (2008). Dissecting GLUT4 traffic components in L6 myocytes by fluorescence-based, single-cell assays. Methods in molecular biology (Clifton, N.J.), 457, 367–378. https://doi.org/10.1007/978-1-59745-261-8_27
Randhawa, V. K., Ishikura, S., Talior-Volodarsky, I., Cheng, A. W., Patel, N., Hartwig, J. H., & Klip, A. (2008). GLUT4 vesicle recruitment and fusion are differentially regulated by Rac, AS160, and Rab8A in muscle cells. The Journal of biological chemistry, 283(40), 27208–27219. https://doi.org/10.1074/jbc.M804282200
Ishikura, S., Antonescu, C. N., & Klip, A. (2010). Documenting GLUT4 exocytosis and endocytosis in muscle cell monolayers. Current protocols in cell biology, Chapter 15, . https://doi.org/10.1002/0471143030.cb1515s46
Chiu, T. T., Sun, Y., Koshkina, A., & Klip, A. (2013). Rac-1 superactivation triggers insulin-independent glucose transporter 4 (GLUT4) translocation that bypasses signaling defects exerted by c-Jun N-terminal kinase (JNK)- and ceramide-induced insulin resistance. The Journal of biological chemistry, 288(24), 17520–17531. https://doi.org/10.1074/jbc.M113.467647
Li, Q., Zhu, X., Ishikura, S., Zhang, D., Gao, J., Sun, Y., Contreras-Ferrat, A., Foley, K. P., Lavandero, S., Yao, Z., Bilan, P. J., Klip, A., & Niu, W. (2014). Ca²⁺ signals promote GLUT4 exocytosis and reduce its endocytosis in muscle cells. American journal of physiology. Endocrinology and metabolism, 307(2), E209–E224. https://doi.org/10.1152/ajpendo.00045.2014
Sun, Y., Jaldin-Fincati, J., Liu, Z., Bilan, P. J., & Klip, A. (2016). A complex of Rab13 with MICAL-L2 and α-actinin-4 is essential for insulin-dependent GLUT4 exocytosis. Molecular biology of the cell, 27(1), 75–89. https://doi.org/10.1091/mbc.E15-05-0319
Jaldin-Fincati, J. R., Bilan, P. J., & Klip, A. (2018). GLUT4 Translocation in Single Muscle Cells in Culture: Epitope Detection by Immunofluorescence. Methods in molecular biology (Clifton, N.J.), 1713, 175–192. https://doi.org/10.1007/978-1-4939-7507-5_14
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