TY - JOUR
T1 - Detection of hepatic maturation by Raman spectroscopy in mesenchymal stromal cells undergoing hepatic differentiation
AU - Wu, Hao Hsiang
AU - Ho, Jennifer H.
AU - Lee, Oscar K.
N1 - Funding Information:
This work was supported in part by the UST-UCSD International Center of Excellence in Advanced Bio-engineering (Grant Number: MOST103-2911-I-009-101), under the Taiwan Ministry of Science and Technology I-RiCE Program. The authors acknowledge financial support from the Ministry of Science and Technology, Taiwan (MOST103-2314-B-010-053-MY3, MOST103-2120-M-010-001, and MOST103-2321-B-010-023), the Ministry of Economic Affairs, Taiwan (103-EC-17-A-17-S1-203), the National Yang-Ming University/ Cheng Hsin General Hospital Grant (CY10405) as well as Wan Fang Hospital/ Taipei Medical University Grant (104swf03). This study was also supported by Aiming for the Top University Plan, a grant from the Ministry of Education. The authors also acknowledge the technical support provided by the Flow cytometry Core Facility of National Yang Ming University and the Imaging Core Facility of Nanotechnology of the UST-YMU.
Publisher Copyright:
© 2016 Wu et al.
PY - 2016/1/11
Y1 - 2016/1/11
N2 - Introduction: 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-1and 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.
AB - Introduction: 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-1and 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.
KW - Hepatic differentiation
KW - Mesenchymal stromal cells
KW - Raman spectroscopy
UR - http://www.scopus.com/inward/record.url?scp=84954179290&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84954179290&partnerID=8YFLogxK
U2 - 10.1186/s13287-015-0259-y
DO - 10.1186/s13287-015-0259-y
M3 - Article
C2 - 26753763
AN - SCOPUS:84954179290
SN - 1757-6512
VL - 7
JO - Stem Cell Research and Therapy
JF - Stem Cell Research and Therapy
IS - 1
M1 - 6
ER -