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

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.

Magnetic levitational bioassembly of 3D tissue construct in space

“…Alternative approaches such as fabricating tissue spheroids from cell monolayers labeled with magnetic nanoparticles ( 46 ) or spontaneous formation of tissue spheroids from monolayer in space ( 40 ) currently do not provide a sufficient number of tissue spheroids with standard size and shape for reproducible experiments. At the same time, the shift to biofabrication of more metabolically active tissue structures, i.e., from epithelial, muscle, or nerve cells, apparently will require the implementation of additional perfusion bioreactors that provide a continuous supply of oxygen and nutrients during the experiment ( 47 , 48 ). However, the application of such systems on ISS requires special equipment and cell culture skills among the astronauts.…”

A Dynamic Culture Method to Produce Ovarian Cancer Spheroids under Physiologically-Relevant Shear Stress

“…Bioreactors are popular tools for large-scale suspension culture of both mammalian and microbial cells, namely for bioproduction [27]. However, the greatest drawback to these systems is the concern about harmful shear stress, namely what is produced by the typically turbulent movement of the stirring apparatus [28]. An example where this apparent weakness is exploited for experimental benefit, however, is with orbital shakers.…”

Design of spherically structured 3D in vitro tumor models -Advances and prospects

“…In fact, 3D spheroids and cellular aggregates obtained in bioreactors frequently exhibit variable shapes and density, leading to inconsistent responses to anti-cancer therapeutics [27,41]. Since cellular concentration is dictated for the entire batch, different internal dynamic flows can also result in aggregates with different sizes [53].…”