Abstract
Metabolic characterization of human adipose tissue-derived mesenchymal stromal/stem cells (ASCs) is of importance in stem cell research. The monitoring of the cell status often requires cell destruction. An analysis of volatile organic compounds (VOCs) in the headspace above cell cultures might be a noninvasive and nondestructive alternative to in vitro analysis. Furthermore, VOC analyses permit new insight into cellular metabolism due to their view on volatile compounds. Therefore, the aim of our study was to compare VOC profiles in the headspace above nondifferentiating and adipogenically differentiating ASCs. To this end, ASCs were cultivated under nondifferentiating and adipogenically differentiating conditions for up to 21 days. At different time points the headspace samples were preconcentrated by needle trap micro extraction and analyzed by gas chromatography/mass spectrometry. Adipogenic differentiation was assessed at equivalent time points. Altogether the emissions of 11 VOCs showed relevant changes and were analyzed in more detail. A few of these VOCs, among them acetaldehyde, were significantly different in the headspace of adipogenically differentiating ASCs and appeared to be linked to metabolic processes. Furthermore, our data indicate that VOC headspace analysis might be a suitable, noninvasive tool for the metabolic monitoring of (mesenchymal stem) cells in vitro.
Highlights
Due to their capacity for self-renewal and multipotent differentiation, mesenchymal stem/stromal cells (MSCs) have been identified as playing an essential role in tissue homeostasis and regeneration [1].In recent years, increasing attention has been paid to MSCs from human adipose tissue, as they show promising potential as a clinical alternative to other MSCs, like bone marrow-derived MSCs [2,3,4]
The assessment of adipogenic differentiation by fluorescent lipid staining revealed that nondifferentiating conditions did not induce lipid accumulation in adipose tissue-derived mesenchymal stromal/stem cells (ASCs) at any time point (Figure 2a,c,e,f), After 24 h, adipogenic stimulation did not lead to lipid accumulation (Figure 2b)
Considerable adipogenic differentiation was clearly visible in almost all cells of the adipogenic cell culture model
Summary
Due to their capacity for self-renewal and multipotent differentiation, mesenchymal stem/stromal cells (MSCs) have been identified as playing an essential role in tissue homeostasis and regeneration [1]. In recent years, increasing attention has been paid to MSCs from human adipose tissue (adipose tissue-derived MSCs, referred to as ASCs), as they show promising potential as a clinical alternative to other MSCs, like bone marrow-derived MSCs (bmMSCs) [2,3,4]. Zuk et al were the first to characterize the mesenchymal differentiation potential of ASCs in more detail [5,6]. Nonmesenchymal differentiation (e.g., neuron-like morphology) of ASCs has been demonstrated [7]. Data concerning the energy metabolism of MSCs in vitro are available but have not been worked out in detail. It has been shown that bmMSCs mainly facilitate glycolysis with subsequent lactate
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