Assembly of three-dimensional polymeric constructs containing cells/biomolecules using carbon dioxide

Using low-pressure carbon dioxide (CO2), we demonstrated a novel and versatile approach to assembling polymeric constructs in the presence of cells and/or biomolecules in an aqueous environment. By regulating the CO2 pressure, the assembly was completed at biologically permissive temperatures with excellent preservation of the original structures. We further demonstrated that mammalian cells can survive the CO2-assisted bioassembly process (37 degrees C, 1.38 MPa, approximately 1 h). Human mesenchymal stem cells from bone marrow (hMSCs) exhibited the same cell morphology and proliferation potential as the untreated control. Mouse embryonic stem cells (mESCs) maintained ES-specific Oct-4 gene expression and differentiation potential after CO2 treatment as well. This method highlights the ability to construct multiple biodegradable polymeric scaffolds with well-defined architecture, on which various types of cells were grown, into a predesigned three-dimensional complex. In addition, protein and DNA bioactivity can be preserved in the context of a CO2-assisted assembly. This CO2-assisted bioassembly method provides for a manufacturing platform that, thus far, has been lacking in the fields of tissue engineering, cell-based biochips, cell therapy, and drug delivery.