title: Detection of hepatic maturation by Raman spectroscopy in mesenchymal stromal cells undergoing hepatic differentiation.
abstract: BACKGROUND
  Mesenchymal stromal cells (MSCs) are well known for their application potential in tissue engineering. We previously reported that MSCs are able to differentiate into hepatocytes in vitro. However, conventional methods for estimating the maturation of hepatic differentiation require relatively large amounts of cell samples. Raman spectroscopy (RS), a photonic tool for acquisition of cell spectra by inelastic scattering, has been recently used as a label-free single-cell detector for biological applications including phenotypic changes and differentiation of cells and diagnosis. In this study, RS is used to real-time monitor the maturation of hepatic differentiation in live MSCs.
  
  
  METHODS
  The MSCs were cultured on the type I collagen pre-coating substrate and differentiated into hepatocytes in vitro using a two-step protocol. The Raman spectra at different time points are acquired in the range 400-3000 cm(-1)and analyzed by quantification methods and principle component analysis during hepatic differentiation from the MSCs.
  
  
  RESULTS
  The intensity of the broad band in the range 2800-3000 cm(-1) reflects the amount of glycogen within lipochrome in differentiated hepatocytes. A high correlation coefficient between the glycogen amount and hepatic maturation was exhibited. Moreover, principle component analysis of the Raman spectra from 400 to 3000 cm(-1) indicated that MSC-derived hepatocytes were close to the primary hepatocytes and were distinct from the undifferentiated MSCs.
  
  
  CONCLUSIONS
  In summary, RS can serve as a rapid, non-invasive, real-time and label-free biosensor and reflects changes in live cell components during hepatic differentiation. The use of RS may thus facilitate the detection of hepatic differentiation and maturation in stem cells. Such an approach may substantially improve the feasibility as well as shorten the time required compared to the conventional molecular biology methods.
authors.affiliation: Institute of Biophotonics, National Yang-Ming University, No. 155, Sec.2, Linong Street, Taipei, 112, Taiwan. haohsiang.wu@gmail.com.
authors.forename: Hao-Hsiang
authors.initials: HH
authors.lastname: Wu
authors.affiliation: Center for Stem Cell Research, Wan Fang Hospital, Taipei Medical University, Taipei, 116, Taiwan. wh9801@yahoo.com.tw.|Graduate Institute of Clinical Medicine, Taipei Medical University, Taipei, Taiwan. wh9801@yahoo.com.tw.|Department of Ophthalmology, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan. wh9801@yahoo.com.tw.
authors.forename: Jennifer H
authors.initials: JH
authors.lastname: Ho
authors.affiliation: Institute of Biophotonics, National Yang-Ming University, No. 155, Sec.2, Linong Street, Taipei, 112, Taiwan. DAV47@tpech.gov.tw.|Taipei City Hospital, No. 145, Zhengzhou Road, Datong District, Taipei, 10341, Taiwan. DAV47@tpech.gov.tw.|Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan. DAV47@tpech.gov.tw.|Stem Cell Research Center, National Yang-Ming University, Taipei, Taiwan. DAV47@tpech.gov.tw.|Department of Medical Research, Taipei Veterans General Hospital, Taipei, Taiwan. DAV47@tpech.gov.tw.|Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan. DAV47@tpech.gov.tw.
authors.forename: Oscar K
authors.initials: OK
authors.lastname: Lee
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D022781:Hepatocytes / Q000502:physiology; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D008807:Mice, Inbred BALB C; D013059:Spectrum Analysis, Raman; D023822:Tissue Engineering