Characterization and Application of a Disposable Rotating Bed Bioreactor for Mesenchymal Stem Cell Expansion.

Maren Loenne,Cornelia Kasper,Alexandra Heilkenbrinker,Anne Neumann,Antonina Lavrentieva

doi:10.3390/bioengineering1040231

Abstract

Recruitment of mesenchymal stromal cells (MSC) into the field of tissue engineering is a promising development since these cells can be expanded vivo to clinically relevant numbers and, after expansion, retain their ability to differentiate into various cell lineages. Safety requirements and the necessity to obtain high cell numbers without frequent subcultivation of cells raised the question of the possibility of expanding MSC in one-way (single-use) disposable bioreactors. In this study, umbilical cord-derived MSC (UC-MSC) were expanded in a disposable Z 2000 H bioreactor under dynamic conditions. Z was characterized regarding residence time and mixing in order to evaluate the optimal bioreactor settings, enabling optimal mass transfer in the absence of shear stress, allowing an reproducible expansion of MSC, while maintaining their stemness properties. Culture of the UC-MSC in disposable Z 2000 H bioreactor resulted in a reproducible 8-fold increase of cell numbers after 5 days. Cells were shown to maintain specific MSC surface marker expression as well as trilineage differentiation potential and lack stress-induced premature senescence.

Highlights

Almost four hundred clinical trials [1] recruiting mesenchymal stem cells (MSC) for the treatment of several diseases such as spinal cord injuries, burns, liver cirrhosis or failure, graft versus host disease, diabetes mellitus, progressive multiple sclerosis and cardiac ischemia are being carried out at the moment
Cultivation in a closed system without the need to manually change the culture medium in a clean bench significantly reduces the risk of possible contamination; Second, the control of all parameters of culture together with continuous documentation using IT systems makes it possible to assure the quality of cells and to avoid human errors; Third, dynamic cultivation systems like bioreactors provide active mass transfer, supplying MSC with gases and nutrients while removing toxic metabolites; Last but not least, there is the argument of production costs - which can be a major obstacle to the spread of MSC clinical applications [6,7,8]
We considered grade of filling of the bioreactor with liquid as well as rotation speed of the reactor bed to have an influence on the residence time and backmixing within the bioreactor

Summary

Introduction

Almost four hundred clinical trials [1] recruiting mesenchymal stem cells (MSC) for the treatment of several diseases such as spinal cord injuries, burns, liver cirrhosis or failure, graft versus host disease, diabetes mellitus, progressive multiple sclerosis and cardiac ischemia are being carried out at the moment. The expansion of MSC in bioreactors is a promising development for the future application of MSC in cell-based therapies and tissue engineering. Cultivation in a closed system without the need to manually change the culture medium in a clean bench significantly reduces the risk of possible contamination; Second, the control of all parameters of culture together with continuous documentation using IT systems makes it possible to assure the quality of cells and to avoid human errors; Third, dynamic cultivation systems like bioreactors provide active mass transfer, supplying MSC with gases and nutrients while removing toxic metabolites; Last but not least, there is the argument of production costs - which can be a major obstacle to the spread of MSC clinical applications [6,7,8]. Several techniques including cultivation on microcarriers, three-dimensional matrix and conventional cell culture plastic surfaces (cell factories) were developed over the past few years

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