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Primary Skeletal Muscle Cells (ATCC® PCS-950-010)

Organism: Homo sapiens, human  /  Tissue: skeletal muscle  / 

Permits and Restrictions

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Organism Homo sapiens, human
Tissue skeletal muscle
Morphology spindle-shaped; elongated (non-differentiated)
Growth Properties adherent
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease normal
Age lot specific
Gender lot-specific
Ethnicity lot-specific
Applications Human Skeletal Muscle Cells (HSkMC) provide an ideal culture model for the study of muscle cell biology, diabetes, insulin receptor studies, muscle cell metabolism, muscle tissue repair, and myotube development.
Product Format frozen 1.0 mL
Storage Conditions liquid nitrogen vapor phase
Comments Characterization: Desmin (+) for differentiated cells, Von Willebrand Factor (-)
Complete Growth Medium

Complete Expansion Medium: One bottle of Mesenchymal Stem Cell Basal Medium (ATCC PCS-500-030) plus one Primary Skeletal Cell Muscle Growth Kit (ATCC PCS-950-040) 

Complete Differentiation Medium: Primary Skeletal Differentiation Tool (ATCC PCS-950-050); a standalone media with no additional supplements required.

Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Subculturing
  1. Passage normal skeletal muscle cells when culture has reached approximately 80% to 90% confluence, and are actively proliferating.
  2. Warm both the Trypsin-EDTA for Primary Cells (ATCC® PCS-999-003) and the Trypsin Neutralizing Solution (ATCC® PCS-999-004) to room temperature prior to dissociation. Warm complete growth medium to 37°C prior to use with the cells.
  3. For each flask, carefully aspirate the spent media without disturbing the monolayer.
  4. Briefly rinse the cell layer with 3 to 5 mL DPBS (ATCC® 30-2200) to remove residual traces of serum and then aspirate and discard the DPBS.
  5. Add pre-warmed trypsin-EDTA solution (2 to 3 mL for every 25 cm2) to each flask.
  6. Gently rock each flask to ensure complete coverage of the trypsin-EDTA solution over the cells.
  7. Observe the cells under the microscope. When the cells pull away from each other and round up (typically within 3 to 5 minutes), remove the flask from the microscope and gently tap it from several sides to promote detachment of the cells from the flask surface.
  8. When the majority of cells are detached, quickly add an equal volume of Trypsin Neutralizing Solution (ATCC® PCS-999-004) to each flask. Gently pipette or swirl the culture to ensure all of the trypsin-EDTA solution has been neutralized.
  9. Transfer the dissociated cells to a sterile centrifuge tube and set aside while processing any remaining cells in the flask.
  10. Add 3 to 5 mL Trypsin Neutralizing Solution to the flask to collect any remaining dissociated cells. Transfer remaining cells into the centrifuge tube.
  11. Repeat steps 10 as needed until all cells have been collected from the flask.
  12. Centrifuge the cells at 150 x g for 3 to 5 minutes.
  13. Carefully aspirate the neutralized dissociation solution from the cell pellet and re-suspend the cells in 5 to 8 mL fresh, pre-warmed, complete growth medium.
  14. Count the cells and seed new flasks at a density of 2,500 to 5,000 cells per cm2.
  15. Place freshly seeded flasks in a 37°C, 5% CO2 incubator for at least 24 to 48 hours before processing the cells further. Refer to Maintenance for guidelines on feeding.
Volume 1.0 mL
Cells per Vial ≥ 5.0 x 105 cells/vial
Sterility Tests Bacteria and yeast: No growth
Mycoplasma: No growth
Viral Testing Hepatitis B: None detected
Hepatitis C: None detected
Human immunodeficiency virus 1: None detected
Human immunodeficiency virus 2: None detected
Population Doubling Time When seeded at 5,000 cells per cm2, cells reach 95% confluence in 4 to 6 days.
C of A
Certificate of Analysis
Certificate of Analysis
Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Hodson N, et al. Characterisation of L-Type Amino Acid Transporter 1 (LAT1) Expression in Human Skeletal Muscle by Immunofluorescent Microscopy. Nutrients 10(1): 1-14, 2017. PubMed: 29278358

Murray IR, et al. Skeletal and cardiac muscle pericytes: Functions and therapeutic potential. Pharmacol Ther 171:65-74, 2017. PubMed: 27595928