Integration of a fiber-based cell culture and biosensing system for monitoring of multiple protein markers secreted from stem cells.

We propose a new platform that can integrate three-dimensional cell culture scaffold and a surface-enhanced Raman spectroscopy (SERS)-based biosensor by stacking them to form a multilayer system, which would allow monitoring of the protein markers secreted from cultured stem cells without periodic cell and/or media collection. The cell culture scaffold supported the proliferation and osteogenic differentiation of adipose-derived mesenchymal stem cells (ADSCs). The SERS capture substrate detected protein markers in combination with SERS tag made with Au-Ag alloy nanoboxes. Incorporating the different Raman reporters into the SERS tag allowed easy identification of target proteins for multiplex assays. The resultant SERS-based immunoassay could detect the pg/mL levels of protein markers without crosstalk and interference. When one ADSC culture scaffold and multiple SERS capture substrates were integrated and incubated in differentiation culture media, our system was sufficiently sensitive to monitor time-dependent secretion of three different osteogenic protein markers from ADSCs during their osteogenic differentiation. Since the sensor and cell culture scaffold can be manipulated independently, various cell and biomarker combinations are possible to obtain relevant information regarding the actual state of the different types of cells.