Raman microspectroscopy: A non-invasive analysis tool for monitoring of collagen-containing extracellular matrix formation in a medium-throughput culture system

A. Kunstar, Cornelis Otto, Hermanus Bernardus Johannes Karperien, Clemens van Blitterswijk, Aart A. van Apeldoorn

Research output: Contribution to journalArticleAcademicpeer-review

15 Citations (Scopus)

Abstract

The three-dimensional environment is known to play an important role in promoting cell–matrix interactions. We have investigated the possibility of using Raman microspectroscopy—which has the great advantage of noninvasive sensing—for in vitro monitoring of extracellular matrix (ECM) formation in a medium-throughput pellet (3D) culture system with soft-litography, agarose-microwell arrays. Chondrocytes were seeded in the agarose microwells in basic or chondrocyte medium. After 3, 7, and 14 days of culture, samples were analyzed for ECM formation by Raman microspectroscopy, histology, and immunofluorescence. ECM formation in the chondrocyte medium-cultured samples was detected by histology and immunofluorescence, and also noninvasively by Raman microspectroscopy. The Raman band of collagen found at 937 cm−1 can be used as a Raman marker for collagen-containing ECM formation over time in the chondrocyte pellets. This culture system can be implemented as a medium-throughput platform for Raman applications and screening microtissue formation, since with these agarose-microwell arrays relatively large numbers of cell pellets could be screened in a short time in situ, without having to transfer the pellets onto microscopic slides. Moreover, in this manner the culture system is suitable for long-term, real-time live-cell measurements.
Original languageEnglish
Pages (from-to)737-744
JournalTissue engineering. Part C: Methods
Volume17
Issue number7
DOIs
Publication statusPublished - 2011

Keywords

  • METIS-282877
  • IR-80639

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