Abstract

With accelerating rates of obesity and type 2 diabetes world-wide, interest in studying the adipocyte and adipose tissue is increasing. Human adipose derived stem cells - differentiated to adipocytes in vitro - are frequently used as a model system for white adipocytes, as most of their pathways and functions resemble mature adipocytes in vivo. However, these cells are not completely like in vivo mature adipocytes. Hosting the cells in a more physiologically relevant environment compared to conventional two-dimensional cell culturing on plastic surfaces, can produce spatial cues that drive the cells towards a more mature state. We investigated the adipogenesis of adipose derived stem cells on electro spun polycaprolactone matrices and compared functionality to conventional two-dimensional cultures as well as to human primary mature adipocytes. To assess the degree of adipogenesis we measured cellular glucose-uptake and lipolysis and used a range of different methods to evaluate lipid accumulation. We compared the averaged results from a whole population with the single cell characteristics – studied by coherent anti-Stokes Raman scattering microscopy - to gain a comprehensive picture of the cell phenotypes. In adipose derived stem cells differentiated on a polycaprolactone-fiber matrix; an increased sensitivity in insulin-stimulated glucose uptake was detected when cells were grown on either aligned or random matrices. Furthermore, comparing differentiation of adipose derived stem cells on aligned polycaprolactone-fiber matrixes, to those differentiated in two-dimensional cultures showed, an increase in the cellular lipid accumulation, and hormone sensitive lipase content. In conclusion, we propose an adipocyte cell model created by differentiation of adipose derived stem cells on aligned polycaprolactone-fiber matrices which demonstrates increased maturity, compared to 2D cultured cells.

Highlights

  • Human adipose derived stem cells are stem cells from the adipose tissue, that proliferate in vitro, and can be stimulated to differentiate into adipocytes [1,2], cartilage [3,4,5], or bone [1,2] cells

  • We aimed to compare the differentiation of human adipose derived stem cells grown as a conventional 2D layer to cells grown on two different electrospun PCL matrices, with either aligned or randomly oriented PCL fibers

  • This study is to our knowledge, the first time Coherent Anti-Stokes Raman Scattering (CARS) and Multi-Photon Excitation Fluorescence (MPEF) microscopy have been combined with a full set of functional measurements of human adipocytes

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Summary

Introduction

Human adipose derived stem cells are stem cells from the adipose tissue, that proliferate in vitro, and can be stimulated to differentiate into adipocytes [1,2], cartilage [3,4,5], or bone [1,2] cells. The morphology of differentiated adipose derived stem cells, with multiple lipid droplets and a comparatively large cytosolic volume, is different to mature adipocytes that have a large central lipid droplet occupying the vast majority of the cell volume. These differences are indicative of the differentiated adipose derived stem cell being an immature model of the adipocyte

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