B). According to the biochemical results, we found that the mechanical properties of the tissue equivalent were affected by culture conditions. After 8 weeks of culture, the shear elastic for modulus (G’) of the tissue equivalent kept under CF did not show any significant variation (from 2.9 x 103 Pa to 3.2 x 103 Pa). Conversely, Inhibitors,research,lifescience,medical under MF

conditions the G’ modulus almost doubled in 8 weeks, jumping from 2.8 x 103 Pa to 5 x 103 Pa. In Figure 2B, the shear elastic modulus at a kinase inhibitor MG132 frequency of 1 Hz has been reported for a biohybrid cultured under MF and CF and compared to that of native dermis. The modulus of the sample cultured under MF is remarkably close to that of native dermis, indicating that a firmer structure has been produced under these processing conditions. Moreover, by comparing the results concerning the biochemical composition and the mechanical properties, it is evident that the higher the accumulation

of GAG and collagen content in the medium, Inhibitors,research,lifescience,medical the lower the modulus of the corresponding biohybrid. Taken together, our results demonstrate the great potential of μTP as functional building blocks in bottom-up tissue engineering. We hypothesized that the added value of using μTP to build up 3D tissue is mainly related to the presence of cells embedded in their own ECM, meaning that cells Inhibitors,research,lifescience,medical undergo μTP assembly with low metabolic output. (or: “meaning that Inhibitors,research,lifescience,medical cells require low metabolic

output to undergo μTP assembly”) Figure 2 Schematic drawing of the culture condition used to culture the μTPs: (A) direct continuous perfusion, CF; (B) tangential nutrient; mixed flow (MF) consists in alternating 1 day perfusion and 1 day tangential flow; (C) GAG and collagen accumulation … Fabrication of Cardiac Inhibitors,research,lifescience,medical Muscle Equivalent In Vitro Cardiac tissue engineering aims to create functional tissue constructs that can serve to re-establish the structure and function of injured myocardium or that can represent in vitro models to study cardiac development and disease. The bottom-up approach previously described to create connective-like tissue in vitro has been used to generate cardiac muscle equivalent (CME). To this end, native cardiac cell population (NCP) seeded into porous gelatine Drug_discovery microbeads was used. The NCP was extracted from neonatal Wistar rats according to the protocol described by Naito et al.28 Once obtained, NCP has been inoculated as full cell population in the spinner flask bioreactor together with porous gelatine microbeads at the ratio of 1,000 cells per bead. Compared with previously described spinner culture conditions used to generate connective-like tissue, the ratio between cell and beads was higher in the cardiac cell culture due to the NCP’s low capability to proliferate in vitro.