GC/MS-based Analysis of Volatile Metabolic Profile Along in vitro Differentiation of Human Induced Pluripotent Stem Cells

Rosamaria Capuano; Rosa Valentina Talarico; Paola Spitalieri; Roberto Paolesse; Giuseppe Novelli; Federica Sangiuolo; Corrado Di Natale

doi:10.21769/BioProtoc.2642

GC/MS-based Analysis of Volatile Metabolic Profile Along in vitro Differentiation of Human Induced Pluripotent Stem Cells

Bio Protoc. 2017 Dec 5;7(23):e2642. doi: 10.21769/BioProtoc.2642.

Authors

Rosamaria Capuano¹, Rosa Valentina Talarico², Paola Spitalieri², Roberto Paolesse³, Giuseppe Novelli², Federica Sangiuolo², Corrado Di Natale²

Affiliations

¹ Department of Electronic Engineering, University of Rome Tor Vergata, Roma, Italy.
² Department of Biomedicine and Prevention, University of Rome Tor Vergata, Rome, Italy.
³ Department of Chemical Science and Technology, University of Rome Tor Vergata, Roma, Italy.

Abstract

Human induced pluripotent stem cells (hiPSCs) are a promising tool in cell-based therapies for degenerative diseases. A safe application of hiPSCs in vivo, requires the detection of the presence of residual undifferentiated pluripotent cells that can potentially cause the insurgence of teratomas. Several studies point out that metabolic products may provide an alternative method to identify the different steps of cells differentiation. In particular, the analysis of volatile organic compounds (VOCs) is gaining a growing interest in this context, thanks to its inherent noninvasiveness. Here, a protocol for VOCs analysis from human induced pluripotent stem cells (hiPSCs) is illustrated. It is based on Solid-Phase Microextraction (SPME) technique coupled with gas chromatography-mass spectrometry (GC/MS). The method is applied to measure the volatile metabolite modifications in cells headspace during cell reprogramming from chorionic villus samples (CVS) to hiPSCs, and along hiPSCs in vitro differentiation into early neural progenitors (NPs), passing through embryoid bodies (EBs) formation.

Keywords: GC/MS; Gas chromatography-mass spectrometry; Human induced pluripotent stem cell; Metabolic profile; SPME; Solid phase microextraction; VOCs; Volatile organic compounds; hiPSC.