Detection of early stage changes associated with adipogenesis using Raman spectroscopy under aseptic conditions

Research output: Contribution to journalArticle

  • External authors:
  • Adam Mitchell
  • Lorna Ashton
  • Xuebin B. Yang
  • Alistair Smith
  • Jennifer Kirkham

Abstract

There is growing interest in the development of methods capable of non-invasive characterization of stem cells prior to their use in cell-based therapies. Raman spectroscopy has previously been used to detect biochemical changes commensurate with the osteogenic, cardiogenic, and neurogenic differentiation of stem cells. The aim of this study was to characterize the adipogenic differentiation of live adipose derived stem cells (ASCs) under aseptic conditions. ASCs were cultured in adipogenic or basal culture medium for 14 days in customized culture flasks containing quartz windows. Raman spectra were acquired every 3 days. Principal component analysis (PCA) was used to identify spectral changes in the cultures over time. Adipogenic differentiation was confirmed using quantitative reverse transcription polymerase chain reaction for the marker genes PPARγ and ADIPOQ and Oil red O staining performed. PCA demonstrated that lipid associated spectral features varied throughout ASC differentiation with the earliest detection of the lipid associated peak at 1,438 cm-1 after 3 days of induction. After 7 days of culture there were clear differences between the spectra acquired from ASCs in adipogenic or basal culture medium. No changes were observed in the spectra acquired from undifferentiated ASCs. Significant up-regulation in the expression of both PPARγ and ADIPOQ genes (P

Bibliographical metadata

Original languageEnglish
Pages (from-to)1012-1019
Number of pages8
JournalCytometry Part A
Volume87
Issue number11
DOIs
StatePublished - 1 Nov 2015