Collagenase digestion was used to isolate subcutaneous and perigonadal cells derived from 6-week old Immortomouse. Cells were grown at +33°C in preadipocyte growth media with 10 units/mL Interferon γ. Aria flow cytometry was used to sort 190 cells and place each one in 190 wells after two passages. Clonal cell lines were created and grown at +33°C with 10 units/ml interferon γ. The clonal cell lines were treated and tested for mycoplasma contamination for 2 weeks. Oil Red O staining was used after differentiation to test the adipogenic capacity of cell lines. Microarray analysis of gene expression is found at the Gene Expression Omnibus database (accession #GSE110531).Subpopulations of adipocytes have distinct cellular responses and metabolism and can be used to study behavioural and physiological differences in fat depots.
Immortalization Method: Conditionally immortalized. Cells were derived from the Immortomouse and grown at +33°C with 10 units/ml interferon-gamma.
BioSafety Level: II
Organism: Mouse (M. musculus)
Species: Mouse
Source Organs: Adipose
Organ Type: Adipose
Growth Properties: Adherent
Morphology: Epithelial-like
Population Doubling: 30-40 hours
Seeding Density: 10,000-20,000 cells/cm
2Markers: Characterization markers (qPCR): Wt1, Lrn4, Upk3b.
Donor Gender: Male
Propagation: The base medium for this cell line is Prigrow III medium available at
abm, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10%, GlutaMAX (Gibco) to a final concentration of 2mM, recombinant mouse IFN-G (Z200085) to a final concentration of 10 U/ml and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.Change media every 2-3 days.Carbon dioxide (CO
2): 5%, Temperature: +33.0°C.* Do not use heat-inactivated FBS for cell culture unless specified otherwise.
Quality Control: 1) qPCR; 2) Oil RedO staining
Shipping Condition: Dry Ice
Storage Condition: liquid nitrogen or -180°C
Reference: Lee, K. Y., Sharma, R., Gase, G., Ussar, S., Li, Y., Welch, L., Berryman, D. E., Kispert, A., Bluher, M., & Kahn, C. R. (2017). Tbx15 Defines a Glycolytic Subpopulation and White Adipocyte Heterogeneity. Diabetes, 66(11), 2822–2829. https://doi.org/10.2337/db17-0218Lee, K. Y., Luong, Q., Sharma, R., Dreyfuss, J. M., Ussar, S., & Kahn, C. R. (2019). Developmental and functional heterogeneity of white adipocytes within a single fat depot. The EMBO journal, 38(3), e99291. https://doi.org/10.15252/embj.201899291