Microbioreactors for Raman Microscopy of Stromal Cell Differentiation

• Published in: Analytical chemistry (Washington) 2010-03, Vol.82 (5), p.1844-1850
• Main Authors: Pully, Vishnu Vardhan, Lenferink, Aufried, van Manen, Henk-Jan, Subramaniam, Vinod, van Blitterswijk, Clemens A, Otto, Cees
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Biological and medical sciences; Bioreactors; Cell culture; Cell Differentiation; Cell differentiation, maturation, development, hematopoiesis; Cell physiology; Chemistry; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Humans; Microscopy - methods; Molecular and cellular biology; Spectrometric and optical methods; Spectrum analysis; Spectrum Analysis, Raman - methods; Stromal Cells - cytology; Tissue engineering
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac902515c

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.