title: 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).
authors: Center for Biosystems Science and Engineering, Indian Institute of Science , Bengaluru, Karnataka, India . ; M D Laboratory, Department of Mechanical Engineering, Indian Institute of Science , Bengaluru, Karnataka, India . ; Saumitra Das's laboratory, Department of Microbiology and Cell Biology, Indian Institute of Science , Bengaluru, Karnataka, India .
authors: Sreenath
authors: S
authors: Balakrishnan
authors: M D Laboratory, Department of Mechanical Engineering, Indian Institute of Science , Bengaluru, Karnataka, India .
authors: M S
authors: MS
authors: Suma
authors: M D Laboratory, Department of Mechanical Engineering, Indian Institute of Science , Bengaluru, Karnataka, India .
authors: Shilpa R
authors: SR
authors: Raju
authors: M D Laboratory, Department of Mechanical Engineering, Indian Institute of Science , Bengaluru, Karnataka, India .
authors: Santosh D B
authors: SD
authors: Bhargav
authors: Saumitra Das's laboratory, Department of Microbiology and Cell Biology, Indian Institute of Science , Bengaluru, Karnataka, India .
authors: S
authors: S
authors: Arunima
authors: Center for Biosystems Science and Engineering, Indian Institute of Science , Bengaluru, Karnataka, India . ; Saumitra Das's laboratory, Department of Microbiology and Cell Biology, Indian Institute of Science , Bengaluru, Karnataka, India .
authors: Saumitra
authors: S
authors: Das
authors: Center for Biosystems Science and Engineering, Indian Institute of Science , Bengaluru, Karnataka, India . ; M D Laboratory, Department of Mechanical Engineering, Indian Institute of Science , Bengaluru, Karnataka, India .
authors: G K
authors: GK
authors: Ananthasuresh
keywords: 3D printing; VeroWhite; bioreactor; live imaging; microfabricated scaffolds; perfusion culture; peristaltic pump