Primary normal human pre-adipocytes (ATCC PCS-210–010, American Type Culture Collection, Manassas, VA, USA) were differentiated into mature adipocytes in wells of 24-well-plates containing adipocyte differentiation medium (Cell Applications, Inc. San Diego, CA) in a 5% CO2 atmosphere at 370 C29 (link). These cells can be expanded in an undifferentiated state for future differentiation to mature adipocytes and show higher efficiency of adipogenesis compared to mesenchymal stem cells. In this study, the cells were cultured in adipocyte differentiation medium with increasing doses of glucose (8.4mM to 19.3mM, Sigma-Aldrich, St. Louis, MO), or ADM (1×10− 10M to 1×10− 8M, Sigma-Aldrich) for 24 hours. Total RNA was isolated from the cells using TRIzol (Life Technologies, Grand Island, NY) and RT was performed for further Quantitative Real-time-PCR analysis.
Atcc pcs210010
Manufactured by American Type Culture Collection
Sourced in United States
The ATCC® PCS210010 is a laboratory equipment product designed for cell culture applications. It provides a controlled environment for the growth and maintenance of cells. The product specifications and core function are available, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Pparγ, by Cell Signaling Technology (2 mentions)
C ebpα, by Cell Signaling Technology (2 mentions)
Gapdh, by Merck Group (2 mentions)
Hmgb2, by Cell Signaling Technology (2 mentions)
Ampkα, by Cell Signaling Technology (2 mentions)
Phospho p ampkα, by Cell Signaling Technology (2 mentions)
P akt1, by Cell Signaling Technology (2 mentions)
L glutamine, by Corning (2 mentions)
Penicillin streptomycin, by Corning (2 mentions)
Dulbecco s modified eagle s medium dmem high glucose, by Corning (2 mentions)
Adipocyte differentiation medium, by Cell Applications (1 mentions)
Adipocyte differentiation medium, by Merck Group (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
4 protocols using atcc pcs210010
1
Adipocyte Differentiation and Glucose/ADM Response
2
Adipocyte Differentiation Protocol
3
Adipocyte Differentiation Protocol
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Mouse embryo fibroblast 3T3-L1 (ATCC®CL173) and normal human primary subcutaneous preadipocytes (ATCC® PCS210010) cells were purchased from American Type Culture Collection (ATCC, Manassas, VA). Dulbecco’s modified Eagle’s medium high glucose (DMEM), penicillin/streptomycin and L-glutamine were purchased from Mediatech, Inc. (Manassas, VA). PPARγ (Cat# 2443S), C/EBPα (Cat# 2295S), FAS (Cat# 4233S), HMGB2 (Cat# 14163S), AMPKα (Cat# 5832S) and phospho(p)-AMPKα (Cat# 50081S-Thr172), Akt1(Cat# 75692S) and p-Akt1 (Cat# 9018S-Ser473) antibodies were from Cell Signaling Technology (Boston, MA). GAPDH (Cat# G9545) antibodies were from Sigma-Aldrich (St. Louis, MO). All secondary antibodies (Cat# 305-035-045) were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA). All other chemicals were obtained from Sigma-Aldrich unless otherwise stated.
4
Differentiation of Primary Human Pre-adipocytes
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Primary normal human pre-adipocytes (ATCC PCS-210-010, American Type Culture Collection, Manassas, VA, USA) were cultured in human preadipocyte growth medium (Cat No. 811-500, Cell Applications, Inc. San Diego, CA) in a 37 °C, 5% CO2 incubator, as previously described29 (link). When cells have reached approximately 80% confluence, the cells were detached using 0.25% trypsin–EDTA (Gibco, Life technology, Gaithersburg, MD), counted, and seeded on 24-well-plates, at a density of 50,000 cells per well in human adipocyte differentiation medium (Cat No. 811D-250, Cell Applications, Inc. San Diego, CA). After 7 days, differentiated adipocytes were used for the total RNA isolation, measurement of mitochondrial contents, assessment of mitochondrial reactive oxygen species (ROS), and the determination of mitochondrial oxygen consumption rate (OCR).
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