The small intestine is a complex tissue with a crypt/villus architecture and high tissue polarity. Maintenance of tissue integrity and function is supported by a constant renewal of the epithelium, with proliferative cells located in the crypts and differentiated cells migrating upward to the top of villi. So far, most in vitro studies have been limited to 2D surfaces or 3D organoid cultures that do not fully recapitulate the tissue 3D architecture, microenvironment and cell compartmentalization found in vivo. Here, we report the development of a 3D model that reproduces more faithfully the architecture of the intestinal epithelium in vitro. We developed a new fabrication process combining a photopolymerizable hydrogel that supports the growth of intestinal cell lines with high-resolution stereolithography 3D printing. This approach offers the possibility to create artificial 3D scaffolds matching the dimensions and architecture of mouse intestinal crypts and villi. We demonstrate that these 3D culture models support the growth and differentiation of Caco-2 cells for 3 weeks. These models may constitute a complementary approach to organoid cultures to study intestinal homeostasis by allowing guided self-organization and controlled differentiation, as well as for in vitro drug screening and testing.
16 February 2018 in Biomedical Physics & Engineering Express
3D fabrication techniques are rapidly expanding in the field of scaffold development for cell culture and tissue engineering. Herein we report the realization of free-standing PEGDA hydrogel architectures by using two-photon lithography. The morphological and immunofluorescence characterization of neuro2A cells revealed a tridimensional colonization featuring multiple neuritic extensions per cell as well as the expression of β -tubulin neuronal marker and actin microfilaments. The results open new perspectives in the continuous quest for structured biomaterials able to provide a favorable environment to cells and at the same time not interfering with imaging protocols necessary for a clear scenario of the cell seeding.
We demonstrate the use of a novel dry film photoresist DF-1000 series for the fabrication of multilevel microfluidic devices by combining a standard lithography technique and lamination technology. The optimization of the technological process enables achievement of high aspect ratio structures: 7 : 1 for free standing structures and 5 : 1 for channel structures. We proved that DF films feature a low autofluorescence level, similar to that of the SU-8 resist and compatible with most lab-on-a-chip applications. The chemical stability against aggressive solvents was also investigated. Last but not least, the non-cytotoxic effect according to ISO 10993-5 on the development of L-929 mouse fibroblast cells was established. Ultimately, we showed that this low-cost material combined with multilevel lamination and UV-lithography techniques allowed the fabrication of 3D microfluidic mixers and opened the way to perform microfluidics in three dimensions.