Approaches for automized expansion and differentiation of human MSC in specialized bioreactors

Anne Neumann,Dominik Egger,Antonina Lavrentieva,Cornelia Kasper,Tim Hatlapatka

doi:10.1186/1753-6561-7-s6-p47

Abstract

Background and experimental approachA main challenge in cell therapies and other tissueregeneration approaches is to produce a therapeuticallysignificant cell number. For expansion of mesenchymalstem cells (MSC) the cultivation on 2D plastic surfacesis still the conventional procedure, even though theculture conditions differ significantly from the 3Denvironment in vivo. Additionally, static amplificationof MSC is a labour-intensive procedure. We thereforeused a specialized rotating bed bioreactor in order tomaximize ex vivo expansion of MSC. MSC were iso-lated from umbilical cord (UC) by explant methodapproach under xeno-free conditions. UC-MSC werethereafter expanded under dynamic conditions in anovel rotating bed bioreactor. The bioreactor systemwas designed to enable integration of sensors for onlinemonitoring of various parameters (e.g. pH, pO

Highlights

Background and experimental approachA main challenge in cell therapies and other tissue regeneration approaches is to produce a therapeutically significant cell number
mesenchymal stem cells (MSC) expanded under dynamic conditions in a rotating bed bioreactor fulfil these MSC criteria
MSC on ceramic materials could be shown to differentiate towards the osteogenic lineage under static conditions

Summary

Introduction

Background and experimental approachA main challenge in cell therapies and other tissue regeneration approaches is to produce a therapeutically significant cell number. For expansion of mesenchymal stem cells (MSC) the cultivation on 2D plastic surfaces is still the conventional procedure, even though the culture conditions differ significantly from the 3D environment in vivo. We used a specialized rotating bed bioreactor in order to maximize ex vivo expansion of MSC. MSC were isolated from umbilical cord (UC) by explant method approach under xeno-free conditions. UC-MSC were thereafter expanded under dynamic conditions in a novel rotating bed bioreactor. Beside cell expansion, directed differentiation of MSC was achieved in bioreactors. MSC lack the ability to grow in 3D direction and build functional tissue in vitro. It is necessary to seed and culture MSC on 3D matrices to obtain functional implants. For guided differentiation towards the osteogenic lineage, MSC were cultivated on ceramic porous matrices under dynamic conditions. Custom-made miniaturized perfusion bioreactors for parallel testing were designed and optimized for that purpose

Methods

Results

Conclusion