A Versatile Bioreactor for Dynamic Suspension Cell Culture. Application to the Culture of Cancer Cell Spheroids

PLoS One. 2016 May 4;11(5):e0154610. doi: 10.1371/journal.pone.0154610. eCollection 2016.

Abstract

A versatile bioreactor suitable for dynamic suspension cell culture under tunable shear stress conditions has been developed and preliminarily tested culturing cancer cell spheroids. By adopting simple technological solutions and avoiding rotating components, the bioreactor exploits the laminar hydrodynamics establishing within the culture chamber enabling dynamic cell suspension in an environment favourable to mass transport, under a wide range of tunable shear stress conditions. The design phase of the device has been supported by multiphysics modelling and has provided a comprehensive analysis of the operating principles of the bioreactor. Moreover, an explanatory example is herein presented with multiphysics simulations used to set the proper bioreactor operating conditions for preliminary in vitro biological tests on a human lung carcinoma cell line. The biological results demonstrate that the ultralow shear dynamic suspension provided by the device is beneficial for culturing cancer cell spheroids. In comparison to the static suspension control, dynamic cell suspension preserves morphological features, promotes intercellular connection, increases spheroid size (2.4-fold increase) and number of cycling cells (1.58-fold increase), and reduces double strand DNA damage (1.5-fold reduction). It is envisioned that the versatility of this bioreactor could allow investigation and expansion of different cell types in the future.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biophysical Phenomena
  • Bioreactors*
  • Cell Culture Techniques / instrumentation*
  • Cell Line, Tumor
  • Computer Simulation
  • Equipment Design
  • Humans
  • Hydrodynamics
  • Microscopy, Electron, Transmission
  • Models, Biological
  • Spheroids, Cellular / cytology*
  • Tumor Cells, Cultured / cytology*

Grants and funding

This work received partial funding from (1) the European FP7 Cooperation - Collaborative Project “Bioactive Highly Porous and Injectable Scaffolds Controlling Stem Cell Recruitment, Proliferation and Differentiation and Enabling Angiogenesis for Cardiovascular Engineered Tissues” - BIOSCENT, 2009-2013 (ID 214539), and (2) the Italian National PRIN 2012 Project “A Bioprocess for the Optimization of 3D Cardiosphere-based constructs for Cardiac Regenerative Medicine” - BEAT3DHEART, 2014-2017 (20123E8FH4), as well as additional funding internal to Politecnico di Torino and Università degli Studi di Parma. Bioexpansys Srl provided financial support in the form of salary for author GFDL and research materials, but had no additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of the authors are articulated in the ‘author contributions’ section.