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iPSC-derived Mesenchymal Stem Cells; BYS0112 (ATCC® ACS-7010)

Organism: Homo sapiens, human  /  Tissue: iPSC-derived Mesenchymal Stem Cells  / 

Permits and Restrictions

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Organism Homo sapiens, human
Tissue iPSC-derived Mesenchymal Stem Cells
Product Format frozen 1.0 mL
Morphology spindle-shaped, fibroblast-like
Culture Properties adherent
Biosafety Level 2

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.

Gender male
Ethnicity Caucasian
Applications Bone cell lineage differentiation, regenerative medicine, cell therapy, exosome research, cancer immunology.
Storage Conditions liquid nitrogen vapor phase
Comments Human iPSC-derived mesenchymal stem cells (MSC) enable researchers to carry out applications that are typically performed using primary MSCs. These cells can be used for bone cell lineage differentiation, regenerative medicine, cell therapy, exosome research, and cancer immunology. 

A distinct advantage of incorporating iPSC-derived MSCs: While there will be lot-to-lot donor variability in primary MSC, there will be no donor variability in iPSC-derived MSC from multiple lots as they are all derived from the parental iPSC line (ATCC ACS-1026). In addition, there is an availability of large number vials of cells from a single donor.

Complete Growth Medium The base medium for these primary cells is Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs (ATCC PCS-500-030). To make the complete medium, add the contents of Mesenchymal Stem Cell Growth Kit for Bone Marrow-derived MSCs (ATCC PCS-500-041) as per the product sheet.
Subculturing
  1. Passage iPSC-derived MSC 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 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. Incubate at 37°C for 3 min. 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 complete growth medium 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 complete growth media 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 270-300 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 in new flasks at a density of 10,000 to 20,000 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.
Cryopreservation Stem Cell Freezing Medium (ATCC ACS-3020)
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial Approximately 2 to 3 x 106 cells
Volume 1.0 mL
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
Viability ≥ 80%
Functional Tests Osteocyte, chondrocyte, and adipocyte differentiation; immunosuppression assay
Population Doubling Time 32 to 35 hours
Name of Depositor ATCC
Year of Origin 2018
References

Zhong H, et al. Human pluripotent stem cell-derived mesenchymal stem cells prevent chronic allergic airway inflammation via TGF-β1-Smad2/Smad3 signaling pathway in mice. Mol Immunol 109: 51-57, 2019. PubMed: 30852246

Jeon OH, et al. Human iPSC-derived osteoblasts and osteoclasts together promote bone regeneration in 3D biomaterials. Sci Rep 6: 26761, 2016. PubMed: 27225733

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
Other Documentation
Restrictions

This product may be used by investigator for research purposes subject to the Limited Use Label License and any additional third party terms. This product is subject to claims under U.S. Patent Nos. 8,058,065 and 8,048,999, pending patent applications, and foreign counterparts thereof.  In addition, this product was generated using the proprietary technology of the DNAVEC Corporation, including, without limitation, recombinant Sendai virus vectors. For information on obtaining additional rights, please contact licensing@atcc.org


References

Zhong H, et al. Human pluripotent stem cell-derived mesenchymal stem cells prevent chronic allergic airway inflammation via TGF-β1-Smad2/Smad3 signaling pathway in mice. Mol Immunol 109: 51-57, 2019. PubMed: 30852246

Jeon OH, et al. Human iPSC-derived osteoblasts and osteoclasts together promote bone regeneration in 3D biomaterials. Sci Rep 6: 26761, 2016. PubMed: 27225733