Controlling Sheer Stress in a Suspension Culture using Couette Flow for Efficient Proliferation of HEK 293 Cells

Katsuhisa Sakaguchi,Tatsuya Shimizu,Azuma Takahashi,Yuji Haraguchi,Mitsuo Umezu,Takanobu Yagi,Sara Suzuki,Nur Khatijah Mohd Zin

doi:10.4172/2476-2296.1000124

Abstract

The suspension culture system is an increasingly popular method of culturing cells not only because of its up scaling ability, but also the non-enzymatic procurement of cells that is crucial for biomedical research, especially in the fields of pharmacology and regenerative medicine. Hypothetically, by controlling and reducing the shear stress applied to cells in a culture system, the higher viability and proliferation rates. In this study, we analyzed HEK 293 cells cultured with a commercially available spinner flask and our newly developed spinner flask which utilizes the theory of Couette flow for controlling shear stress. Fluid analysis and metabolic analysis of the cultured cells were measured at three different rotational speeds, 40, 50 and 60 rpm. It was apparent that 50 rpm was by far the best speed to proliferate the cells. A further viability test was also done in order to validate our hypothesis. Furthermore, by using the metabolic analysis results, it was observed that in the controlled stress system, the consumption of glucose doubled and lactate production was significantly higher compared to cells that were maintained in the conventional suspension method. Thus, Couette flow based suspension culture system will be a major contributor to the future biomedical and pharmacological field.

Highlights

Biopharmaceutical products such as hormones, antibody preparations, blood factors, cytokines, growth factors and vaccines have become a major interest worldwide [1]
Mammalian cell culture has come to the forefront of biopharmaceutical production due to its capability of properly folding, assembly, and post-translational modification (PTM) of proteins [2]
Industrially produced recombinant proteins from mammalian cells are used in a wide variety of biomedical research at laboratories around the world

Summary

Introduction

Biopharmaceutical products such as hormones, antibody preparations, blood factors, cytokines, growth factors and vaccines have become a major interest worldwide [1]. The idea of controlling shear stress and utilizing a three-dimensional (3-D) printer, this idea was tested using HEK 293 cells that were proliferated efficiently and had a higher viability than those from a conventional system. Fluid dynamical analysis The fluid movement in both the conventional and controlled shear stress methods of the suspension system was recorded and analyzed as shown in Figures 2a, 2b and 2c.

Results

Conclusion