Adipogenesis of murine embryonic stem cells in a three-dimensional culture system using electrospun polymer scaffolds.
A mechanistic understanding of adipose tissue differentiation is critical for the treatment and prevention of obesity and type 2 diabetes. Conventional in vitro models of adipogenesis are preadipocytes or freshly isolated adipocytes grown in two-dimensional (2D) cultures. Optimal results using in vitro tissue culture models can be expected only when adipocyte models closely resemble adipose tissue in vivo. Thus the design of an in vitro three-dimensional (3D) model which faithfully mimics the in vivo environment is needed to effectively study adipogenesis. Pluripotent embryonic stem (ES) cells are a self-renewing cell type that can readily be differentiated into adipocytes. In this study, a 3D culture system was developed to mimic the geometry of adipose tissue in vivo. Murine ES cells were seeded into electrospun polycaprolactone scaffolds and differentiated into adipocytes in situ by hormone induction as demonstrated using a battery of gene and protein expression markers along with the accumulation of neutral lipid droplets. Insulin-responsive Akt phosphorylation, and beta-adrenergic stimulation of cyclic AMP synthesis were demonstrated in ES cell-derived adipocytes. Morphologically, ES cell-derived adipocytes resembled native fat cells by scanning electron and phase contrast microscopy. This tissue engineered ES cell-matrix model has potential uses in drug screening and other therapeutic developments.
Kang
Xihai
X
Department of Obstetrics and Gynecology (Laboratory of Perinatal Research), The Ohio State University, College of Medicine, 1654 Upham Drive, Means Hall, Fifth Floor, Columbus, OH 43210, USA.
Xie
Yubing
Y
Powell
Heather M
HM
James Lee
L
L
Belury
Martha A
MA
Lannutti
John J
JJ
Kniss
Douglas A
DA
D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism; D050156:Adipogenesis; D000273:Adipose Tissue / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D004563:Electrochemistry / Q000379:methods; D004622:Embryo, Mammalian / Q000166:cytology*; D007328:Insulin / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D051379:Mice; D010766:Phosphorylation; D011108:Polymers / Q000737:chemistry*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology*
D007328:Insulin; D008055:Lipids; D011108:Polymers; D051057:Proto-Oncogene Proteins c-akt