Customizable 3D-printed (co-)cultivation systems for in vitro study of angiogenesis

Due to the ever-increasing resolution of 3D printing technology, additive manufacturing is now even used to produce complex devices for laboratory applications. Personalized experimental devices or entire cultivation systems of almost unlimited complexity can potentially be manufactured within hours from start to finish—an enormous potential for experimental parallelization in a highly controllable environment. This study presents customized 3D-printed co-cultivation systems, which qualify for angiogenesis studies. In these systems, endothelial and mesenchymal stem cells (AD-MSC) were indirectly co-cultivated—that is, both cell types were physically separated through a rigid, 3D-printed barrier in the middle, while still sharing the same cell culture medium that allows for the exchange of signalling molecules. Biochemical-based cytotoxicity assays initially confirmed that the 3D printing material does not exert any negative effects on cells. Since the material also enables phase contrast and fluorescence microscopy, the behaviour of cells could be observed over the entire cultivation via both. Microscopic observations and subsequent quantitative analysis revealed that endothelial cells form tubular-like structures as angiogenic feature when indirectly co-cultured alongside AD-MSCs in the 3D-printed co-cultivation system. In addition, further 3D-printed devices are also introduced that address different issues and aspire to help in varying experimental setups. Our results mark an important step forward for the integration of customized 3D-printed systems as self-contained test systems or equipment in biomedical applications. © 2020 by the authors. Licensee MDPI, Basel, Switzerland

3D-printed flow cells for aptamer-based impedimetric detection of e. coli crooks strain

Electrochemical spectroscopy enables rapid, sensitive, and label-free analyte detection without the need of extensive and laborious labeling procedures and sample preparation. In addition, with the emergence of commercially available screen-printed electrodes (SPEs), a valuable, disposable alternative to costly bulk electrodes for electrochemical (bio-)sensor applications was established in recent years. However, applications with bare SPEs are limited and many applications demand additional/supporting structures or flow cells. Here, high-resolution 3D printing technology presents an ideal tool for the rapid and flexible fabrication of tailor-made, experiment-specific systems. In this work, flow cells for SPE-based electrochemical (bio-)sensor applications were designed and 3D printed. The successful implementation was demonstrated in an aptamer-based impedimetric biosensor approach for the detection of Escherichia coli (E. coli) Crooks strain as a proof of concept. Moreover, further developments towards a 3D-printed microfluidic flow cell with an integrated micromixer also illustrate the great potential of high-resolution 3D printing technology to enable homogeneous mixing of reagents or sample solutions in (bio-)sensor applications

Real-time live-cell imaging technology enables high-throughput screening to verify in vitro biocompatibility of 3D printed materials

With growing advances in three-dimensional (3D) printing technology, the availability and diversity of printing materials has rapidly increased over the last years. 3D printing has quickly become a useful tool for biomedical and various laboratory applications, offering a tremendous potential for efficiently fabricating complex devices in a short period of time. However, there still remains a lack of information regarding the impact of printing materials and post-processing techniques on cell behavior. This study introduces real-time live-cell imaging technology as a fast, user-friendly, and high-throughput screening strategy to verify the in vitro biocompatibility of 3D printed materials. Polyacrylate-based photopolymer material was printed using high-resolution 3D printing techniques, post-processed using three different procedures, and then analyzed with respect to its effects on cell viability, apoptosis, and necrosis of adipogenic mesenchymal stem cells (MSCs). When using ethanol for the post-processing procedure and disinfection, no significant effects on MSCs could be detected. For the analyses a novel image-based live-cell analysis system was compared against a biochemical-based standard plate reader assay and traditional flow cytometry. This comparison illustrates the superiority of using image-based detection of in vitro biocompatibility with respect to analysis time, usability, and scientific outcome

3D Printed Microfluidic Mixers—A Comparative Study on Mixing Unit Performances

One of the basic operations in microfluidic systems for biological and chemical applications is the rapid mixing of different fluids. However, flow profiles in microfluidic systems are laminar, which means molecular diffusion is the only mixing effect. Therefore, mixing structures are crucial to enable more efficient mixing in shorter times. Since traditional microfabrication methods remain laborious and expensive, 3D printing has emerged as a potential alternative for the fabrication of microfluidic devices. In this work, five different passive micromixers known from literature are redesigned in comparable dimensions and manufactured using high‐definition MultiJet 3D printing. Their mixing performance is evaluated experimentally, using sodium hydroxide and phenolphthalein solutions, and numerically via computational fluid dynamics. Both experimental and numerical analysis results show that HC and Tesla‐like mixers achieve complete mixing after 0.99 s and 0.78 s, respectively, at the highest flow rate (Reynolds number (Re) = 37.04). In comparison, Caterpillar mixers exhibit a lower mixing rate with complete mixing after 1.46 s and 1.9 s. Furthermore, the HC mixer achieves very good mixing performances over all flow rates (Re = 3.7 to 37.04), while other mixers show improved mixing only at higher flow rates

3D Printed Microfluidic Mixers—A Comparative Study on Mixing Unit Performances

One of the basic operations in microfluidic systems for biological and chemical applications is the rapid mixing of different fluids. However, flow profiles in microfluidic systems are laminar, which means molecular diffusion is the only mixing effect. Therefore, mixing structures are crucial to enable more efficient mixing in shorter times. Since traditional microfabrication methods remain laborious and expensive, 3D printing has emerged as a potential alternative for the fabrication of microfluidic devices. In this work, five different passive micromixers known from literature are redesigned in comparable dimensions and manufactured using high‐definition MultiJet 3D printing. Their mixing performance is evaluated experimentally, using sodium hydroxide and phenolphthalein solutions, and numerically via computational fluid dynamics. Both experimental and numerical analysis results show that HC and Tesla‐like mixers achieve complete mixing after 0.99 s and 0.78 s, respectively, at the highest flow rate (Reynolds number (Re) = 37.04). In comparison, Caterpillar mixers exhibit a lower mixing rate with complete mixing after 1.46 s and 1.9 s. Furthermore, the HC mixer achieves very good mixing performances over all flow rates (Re = 3.7 to 37.04), while other mixers show improved mixing only at higher flow rates

Congreso Internacional de Bioeconomía Circular - Fase III 3/3

Moderador: Dr. José de Jesús Brambila (COLPOS) Dr. Salvador Peniche Camps, Lic. Diana Stefania García Valadez, U. de G. Mtra. Carmen Girón Domínguez, Fundación CTA, España Dr. Jorge H. Siller Cepeda, Simplicidad y Enfoque Sostenible Dra. Magdalena Rojas Rojas, UACH Dra. Karina Valencia Sandoval, UAEH Dra. Alejandra Corichi García, UAEH Mtra. Ana Karen Miranda, UACH Dr. Enrique Mendoza Tello, Síntesis AC Mtra Ina Daniela Maza Villalobos Dr. Salvador Arturo Velázquez Crôtte, U. de G.-CUCEA México Moderador: Dr. Mario del Roble Pensado Leglise, IPN CIIEMAD Dra. Miriam Edith García Salazar, Cátedras CONACYT Dr. Sergio Gabriel Ceballos Pérez, COLPOS Psnt. Edgar Corona Zamora, UNAM Psnt. Cristal Antúnez, UNAM Dra. María del Rosario Reyes, ECOSUR Dr. Carlos R. Menéndez G. IICA México M.C. Verónica Estela Ruiz, UACH Ciencias Agrarias Mtra. Nelly López Azuz, IIA - UNAM / México M.C. Carlos Mallén Rivera, INIFAP/México Dr. Pedro Gutiérrez Yurrita, Profepa/MéxicoTemas a tratar: Los senderos de la bioeconomía, logros y retos científicos, institucionales y de gobernanza