A Scalable Perfusion Culture System with Miniature Peristaltic Pumps for Live-Cell Imaging Assays with Provision for Microfabricated Scaffolds

Abstract

We present a perfusion culture system with miniature bioreactors and peristaltic pumps. The bioreactors are designed for perfusion, live-cell imaging studies, easy incorporation of microfabricated scaffolds, and convenience of operation in standard cell culture techniques. By combining with miniature peristaltic pumps—one for each bioreactor to avoid cross-contamination and to maintain desired flow rate in each—we have made a culture system that facilitates perfusion culture inside standard incubators. This scalable system can support multiple parallel perfusion experiments. The major components are fabricated by three-dimensional printing using VeroWhite, which we show to be amenable to ex vivo cell culture. Furthermore, the components of the system can be reused, thus making it economical. We validate the system and illustrate its versatility by culturing primary rat hepatocytes, live imaging the growth of mouse fibroblasts (NIH 3T3) on microfabricated ring-scaffolds inserted into the bioreactor, performing perfusion culture of breast cancer cells (MCF7), and high-magnification imaging of hepatocarcinoma cells (HuH7).

Feasibility and Biocompatibility of 3D‐Printed Photopolymerized and Laser Sintered Polymers for Neuronal, Myogenic, and Hepatic Cell Types

“…Such responses may present the opportunity to utilize these materials within devices in which cell/polymer interaction is confined to leachates within the culture medium. Pertinent to this are bespoke perfusion systems and microfluidic devices that are currently being utilized for the examination and recreation of musculoskeletal and neurological cellular physiological processes in vitro . The homogeneity of cytochrome P450 enzymatic activity across SL, LS, and PolyJet AM processes may also afford the opportunity to produce 3D‐printed fluidic devices that contain multiple cell types ( Table 2 ).…”

“…The possibility of 3D printing (also known as additive manufacturing (AM)) bespoke biologically receptive parts, has driven the increasing application of AM technologies in biological systems. Techniques such as fused deposition modeling (FDM), photopolymerization processes such as stereolithography (SL) and PolyJet printing, in addition to powder‐based particle consolidation laser sintering (LS) technology have all previously been utilized to provide devices/scaffolds for bioengineering . However, the numerous advantages of AM, including design freedom and rapid production without molds or tooling, are currently offset by a lack of fully characterized biocompatible materials …”

Biocompatible 3D printed polymers via fused deposition modelling direct C2C12 cellular phenotype in vitro

“…This may hold specific relevance within flow reactor applications of 3D printing, where it is of paramount importance to understand the effects of polymer chemical leachate on cellular behaviour, with many parts designed where polymer interaction is often confined to perfused medium, such as live-cell imaging systems. 2 Differential myotube phenotypes were observed in cells cultured on 3D printed polymers, with reductions in width documented in PLA, PET and PC conditions (Fig. 5A).…”

Enhancing cell and gene therapy manufacture through the application of advanced fluorescent optical sensors (Review)

“…Therefore, as the volume of the bioreactors increase the need for sensory input also increases, as they are prone to the development of dead-spots, which are regions with suboptimal gas and media perfusion. 16 Examples of critical quality attributes (CQAs) that would benefit from sensors and enhance bioreactor performance include: small molecules and ions (pH, oxygen, and carbon dioxide), metabolites (glucose, lactate, and ammonia), and large macromolecules (enzymes, growth factors, and cytokines). [17][18][19] B.…”