L6 Rat Myoblast Cell Line

Immortalized rat skeletal (L6) myoblast cell line that was selected for high fusion potential and endogenous expression of GLUT4 in the myotube stage.

Highlights:

  • Differentiate with high reliability into a myotube muscle cell phenotype that naturally expresses the GLUT4 glucose transporter protein
  • Significant insulin-stimulated glucose uptake biological response
  • Useful for novel anti-diabetic compound screening
  • Amendable for transient transfection by plasmid-based gene transfer and viral infection protocols (retrovirus and adenovirus)

Insulin stimulation of glucose transport in skeletal muscle results mainly due to translocation of the glucose transporter GLUT4 to the cell surface. L6 cells, originally derived from rat skeletal muscle, propagate as mononucleated myoblasts but can differentiate into multinucleated primary myotubes. The myotubes express several proteins typical of skeletal muscle including the GLUT4 glucose transporter. Insulin stimulates glucose uptake with high sensitivity and maximal responsiveness only in differentiated L6 myotubes and GLUT4 expression parallels the acquisition of these characteristics as the L6 cells differentiate. These features of L6 myotubes are important since GLUT4 is responsible for insulin-dependent glucose uptake in mature skeletal muscle.

Also available: L6-GLUT4myc Rat Myoblast Cell Line

From the laboratory of Amira Klip, PhD, Hospital For Sick Children.

Catalog Number Product DataSheet Size AVAILABILITY Price Qty
ESK201-FP
L6 Rat Myoblast Cell Line
1 vial In stock
Regular Price:$519.00
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Specifications

Product Type: Cell Line
Name: L6
Cell Type: Skeletal muscle
Organism: Rat
Accession ID: CVCL_0385
Source: Quadriceps
Morphology: Myoblast/Myotube
Biosafety Level: II
Subculturing: 2-3 days
Growth Conditions: MEM-α (with Ribonucleosides and Deoxyribonucleosides) +10% FBS (See protocol below)
Cryopreservation: MEM-α +10% FBS+ 10% DMSO
Storage: Liquid nitrogen
Shipped: Dry ice

Documentation
Provider
From the laboratory of Amira Klip, PhD, Hospital For Sick Children.
Comments

Myoblasts should be passaged at approximately 70% of confluence. Myoblasts can be differentiated into myotubes by switching confluent myoblast monolayers to MEM-a +2% FBS for 4-5 days. Upon differentiation, the myotubes express more skeletal muscle proteins and based on our research, the GLUT4 glucose transporter, which is necessary for the development of insulin-stimulated glucose uptake. The insulin responsiveness of transporter-facilitated glucose uptake is ~2-fold in the myotube culture.

References

Cell Line References

  1. Mitsumoto Y, Burdett E, Grant A, Klip A. (1991) Differential expression of the GLUT1 and GLUT4 glucose transporters during differentiation of L6 muscle cells. Biochem Biophys Res Commun. 175: 652-9.
  2. Tsakiridis T, Vranic M, Klip A. (1994) Disassembly of the Actin Network Inhibits Insulin-dependent Stimulation of Glucose Transport and Prevents Recruitment of Glucose Transporters to the Plasma Membrane. J. Biol. Chem. 269: 29934-42.
  3. Huang C, Somwar R, Patel N, Niu W, Török D, Klip A. Sustained exposure of L6 myotubes to high glucose and insulin decreases insulin-stimulated GLUT4 translocation but upregulates GLUT4 activity. Diabetes. 2002 Jul;51(7):2090-8.

Application References

  1. Pillon NJ, Li YE, Fink LN, Brozinick JT, Nikolayev A, Kuo MS, Bilan PJ, KlipA. (2014) Nucleotides released from palmitate-challenged muscle cells through pannexin-3 attract monocytes. Diabetes. 63: 3815-26.
  2. Li Q, Zhu X, Ishikura S, Zhang D, Gao J, Sun Y, Contreras-Ferrat A, Foley KP, Lavandero S, Yao Z, Bilan PJ, Klip A, Niu W. (2014) Ca²? signals promote GLUT4 exocytosis and reduce its endocytosis in muscle cells. Am J Physiol Endocrinol Metab. Jul 15;307(2):E209-24.
  3. Sun Y, Chiu TT, Foley KP, Bilan PJ, Klip A. (2014) Myosin Va mediates Rab8A-regulated GLUT4 vesicle exocytosis in insulin-stimulated muscle cells. Mol Biol Cell. 25: 1159-70.
  4. Foley KP, Klip A. (2014) Dynamic GLUT4 sorting through a syntaxin-6 compartment in muscle cells is derailed by insulin resistance-causing ceramide. Biol Open. 3: 314-25.
  5. Chiu TT, 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. J Biol Chem. 288:17520-31.
  6. Pillon NJ, Arane K, Bilan PJ, Chiu TT, Klip A. (2012) Muscle cells challenged with saturated fatty acids mount an autonomous inflammatory response that activates macrophages. Cell Commun Signal. 10: 30.
  7. Boguslavsky S, Chiu T, Foley KP, Osorio-Fuentealba C, Antonescu CN, Bayer KU, Bilan PJ, Klip A. (2012) Myo1c binding to submembrane actin mediates insulin-induced tethering of GLUT4 vesicles. Mol Biol Cell. 23: 4065-78.
  8. Kewalramani G, Fink LN, Asadi F, Klip A. (2011) Palmitate-activated macrophages confer insulin resistance to muscle cells by a mechanism involving protein kinase C ? and ?. PLoS One. 6: e26947.
  9. Sun Y, Bilan PJ, Liu Z, Klip A. (2010) Rab8A and Rab13 are activated by insulin and regulate GLUT4 translocation in muscle cells. Proc Natl Acad Sci U S A. 107: 19909-14.
  10. Ishikura S, Antonescu CN, Klip A. (2010) Documenting GLUT4 Exocytosis and Endocytosis in Muscle Cell Monolayers. Curr. Protoc. Cell Biol., John Wiley & Sons, Inc., 46: Unit 15.15: 1-9.
  11. Samokhvalov V, Bilan PJ, Schertzer JD, Antonescu CN, Klip A. (2009) Palmitate- and lipopolysaccharide-activated macrophages evoke contrasting insulin responses in muscle cells. Am J Physiol Endocrinol Metab. 296: E37-46.
  12. Antonescu CN, Díaz M, Femia G, Planas JV, Klip A. (2008) Clathrin-dependent and independent endocytosis of glucose transporter 4 (GLUT4) in myoblasts: regulation by mitochondrial uncoupling. Traffic. 9: 1173-90.
  13. Thong FS, Bilan PJ, Klip A. (2007) The Rab GTPase-activating protein AS160 integrates Akt, protein kinase C, and AMP-activated protein kinase signals regulating GLUT4 traffic. Diabetes. 56: 414-23.
  14. Wijesekara N, Tung A, Thong F, Klip A. (2006) Muscle cell depolarization induces again in surface GLUT4 via reduced endocytosis independently of AMPK. Am J Physiol Endocrinol Metab. 290: E1276-86.
  15. Ishiki M, Randhawa VK, Poon V, Jebailey L, Klip A. (2005) Insulin regulates the membrane arrival, fusion, and C-terminal unmasking of glucose transporter-4 via distinct phosphoinositides. J Biol Chem. 280: 28792-802.
  16. Rudich A, Konrad D, Török D, Ben-Romano R, Huang C, Niu W, Garg RR, Wijesekara N, Germinario RJ, Bilan PJ, Klip A. (2003) Indinavir uncovers different contributions of GLUT4 and GLUT1 towards glucose uptake in muscle and fat cells and tissues. Diabetologia 46: 649-58.
  17. Wang Q, Somwar R, Bilan PJ, Liu Z, Jin J, Woodgett JR, Klip A. (1999) Protein kinase B/Akt participates in GLUT4 translocation by insulin in L6 myoblasts. Mol Cell Biol. 19: 4008-18.
  18. 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: 193-7.
  19. Jaldin-Fincati JR, Bilan PJ, Klip A. GLUT4 Translocation in Single Muscle Cells in Culture: Epitope Detection by Immunofluorescence. Methods Mol Biol. 2018;1713:175-192. View Article
  20. Kalinovich A, Dehvari N, Åslund A, et al. Treatment with a β-2-adrenoceptor agonist stimulates glucose uptake in skeletal muscle and improves glucose homeostasis, insulin resistance and hepatic steatosis in mice with diet-induced obesity [published online ahead of print, 2020 May 29]. Diabetologia. 2020;10.1007/s00125-020-05171-y. View Article
  21. Kalinovich A, Dehvari N, Åslund A, et al. Treatment with a β-2-adrenoceptor agonist stimulates glucose uptake in skeletal muscle and improves glucose homeostasis, insulin resistance and hepatic steatosis in mice with diet-induced obesity. Diabetologia. 2020;63(8):1603-1615. View Article

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