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Turner PA, Gurumurthy B, Bailey JL, et al. Adipogenic Differentiation of Human Adipose-Derived Stem Cells Grown as Spheroids. Process Biochem. 2017;59:312-320doi: 10.1016/j.procbio.2017.02.003.
Turner, P. A., Gurumurthy, B., Bailey, J. L., Elks, C. M., & Janorkar, A. V. (2017). Adipogenic Differentiation of Human Adipose-Derived Stem Cells Grown as Spheroids. Process biochemistry (Barking, London, England), 59312-320. https://doi.org/10.1016/j.procbio.2017.02.003
Turner, Paul A, et al. "Adipogenic Differentiation of Human Adipose-Derived Stem Cells Grown as Spheroids." Process biochemistry (Barking, London, England) vol. 59 (2017): 312-320. doi: https://doi.org/10.1016/j.procbio.2017.02.003
Turner PA, Gurumurthy B, Bailey JL, Elks CM, Janorkar AV. Adipogenic Differentiation of Human Adipose-Derived Stem Cells Grown as Spheroids. Process Biochem. 2017 Aug;59:312-320. doi: 10.1016/j.procbio.2017.02.003. Epub 2017 Feb 06. PMID: 28966553; PMCID: PMC5617119.
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Process Biochem. 2017 Aug;59:312-320. doi: 10.1016/j.procbio.2017.02.003. Epub 2017 Feb 06.

Adipogenic Differentiation of Human Adipose-Derived Stem Cells Grown as Spheroids.

Process biochemistry (Barking, London, England)

Paul A Turner, Bhuvaneswari Gurumurthy, Jennifer L Bailey, Carrie M Elks, Amol V Janorkar

Affiliations

  1. Department of Biomedical Materials Science, School of Dentistry, University of Mississippi Medical Center, 2500 N State Street, Jackson, MS.
  2. Matrix Biology Laboratory, Pennington Biomedical Research Center, 6400 Perkins Road, Baton Rouge, LA.

PMID: 28966553 PMCID: PMC5617119 DOI: 10.1016/j.procbio.2017.02.003

Abstract

Understanding the process of adipogenesis is critical if suitable therapeutics for obesity and related metabolic diseases are to be found. The current study presents proof of feasibility of creating a 3-D spheroid model using human adipose-derived stem cells (hASCs) and their subsequent adipogenic differentiation. hASC spheroids were formed atop an elastin-like polypeptide-polyethyleneimine (ELP-PEI) surface and differentiated using an adipogenic cocktail. Spheroids were matured in the presence of dietary fatty acids (linoleic or oleic acid) and evaluated based on functional markers including intracellular protein, CD36 expression, triglyceride accumulation, and PPAR-γ gene expression. Spheroid size was found to increase as the hASCs matured in the adipocyte maintenance medium, though the fatty acid treatment generally resulted in smaller spheroids compared to control. A stable protein content over the 10-day maturation period indicated contact-inhibited proliferation as well as minimal loss of spheroids during culture. Spheroids treated with fatty acids showed greater amounts of intracellular triglyceride content and greater expression of the key adipogenic gene, PPAR-γ. We also demonstrated that 3-D spheroids outperformed 2-D monolayer cultures in adipogenesis. We then compared the adipogenesis of hASC spheroids to that in 3T3-L1 spheroids and found that the triglyceride accumulation was less profound in hASC spheroids than that in 3T3-L1 adipocytes, correlated with smaller average spheroids, suggesting a relatively slower differentiation process. Taken together, we have shown the feasibility of adipogenic differentiation of patient-derived hASC spheroids, which with further development, may help elucidate key features in the adipogenesis process.

Keywords: 3-D cell culture; adipose tissue model; cell morphology; elastin-like polypeptide; spheroids; stem cells

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