Microbioreactors for Raman microscopy of stromal cell differentiation.
We present the development of microbioreactors with a sensitive and accurate optical coupling to a confocal Raman microspectrometer. We show that such devices enable in situ and in vitro investigation of cell cultures for tissue engineering by chemically sensitive Raman spectroscopic imaging techniques. The optical resolution of the Raman microspectrometer allows recognition and chemical analysis of subcellular features. Human bone marrow stromal cells (hBMSCs) have been followed after seeding through a phase of early proliferation until typically 21 days later, well after the cells have differentiated to osteoblasts. Long-term perfusion of cells in the dynamic culture conditions was shown to be compatible with experimental optical demands and off-line optical analysis. We show that Raman optical analysis of cells and cellular differentiation in microbioreactors is feasible down to the level of subcellular organelles during development. We conclude that microbioreactors combined with Raman microspectroscopy are a valuable tool to study hBMSC proliferation, differentiation, and development into tissues under in situ and in vitro conditions.
Biophysical Engineering Group, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, 7522 ND Enschede, The Netherlands.
Vishnu Vardhan
VV
Pully
Aufried
A
Lenferink
Henk-Jan
HJ
van Manen
Vinod
V
Subramaniam
Clemens A
CA
van Blitterswijk
Cees
C
Otto
D019149:Bioreactors; D002454:Cell Differentiation; D006801:Humans; D008853:Microscopy / Q000379:methods*; D013059:Spectrum Analysis, Raman / Q000379:methods*; D017154:Stromal Cells / Q000166:cytology*