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POSTER PRESENTATION Open Access Expansion of mesenchymal adipose-tissue derived stem cells in a stirred single-use bioreactor under low-serum conditions Carmen Schirmaier 1* , Stephan C Kaiser 1 , Valentin Jossen 1 , Silke Brill 2 , Frank Jüngerkes 2 , Christian van den Bos 2 , Dieter Eibl 1 , Regine Eibl 1 From 23rd European Society for Animal Cell Technology (ESACT) Meeting: Better Cells for Better Health Lille, France. 23-26 June 2013 Background The need for human mesenchymal stem cells (hMSCs) has increased enormously in recent years due to their important therapeutic potential. Efficient cell expansion is essential to providing clinically relevant cell numbers. Such cell quantities can be manufactured by means of scalable microcarrier (MC)-supported cultivations in stirred single-use bioreactors. Materials and methods Preliminary tests in disposable-spinners (100 mL culture volume, Corning) were used to determine two suitable media and MC-types for serum reduced expansions (< 10%) of human adipose tissue-derived stem cells (hADSCs; passage 2, Lonza). Using such optimized media-MC-combinations, hADSCs expanded 30 to 40- fold, which compares well with expansion rates in planar culture. Based on computational fluid dynamics simula- tions and suspension analyses in spinners [1], optimal operating parameters were determined in a BIOSTAT ® UniVessel ® SU 2 L (2 L culture volume, Sartorius Stedim Biotech). Results In subsequent batch tests with the BIOSTAT UniVessel ® SU 2 L, expansion rates of between 30 and 40-fold were reached and up to 4.4·10 8 cells with a cell viability exceeding 98% were harvested. Flow cytometry tests demonstrated typical marker profiles following cell expansion and harvest. A 40-fold expansion rate delivered a total of 1·10 10 cells in a first cultivation with the BIOSTAT ® CultiBag STR 50 L (35 L culture volume, Sartorius Stedim Biotech). Conclusions In summary, the foundations for successfully expanding therapeutic stem cells in truly scalable systems have been laid. Strategies ensuring expansion rates between 60 and 70-fold and, thus, generating cell amounts over 10 10 are now in preparation. Acknowledgements This work is part of the project Development of a technology platform for a scalable production of therapeutically relevant stem cells(No. 12893.1 VOUCH-LS). It is supported by the Commission for Technology and Innovation (CTI, Switzerland). The authors would like to thank the CTI for partially financing the investigations presented. Authorsdetails 1 Zurich University of Applied Sciences, Institute of Biotechnology, Biochemical Engineering and Cell Cultivation Technique, 8820 Wädenswil, Switzerland. 2 Lonza Cologne GmbH, 50829 Cologne, Germany. Published: 4 December 2013 Reference 1. Kaiser S C, Jossen V, Schirmaier C, Eibl D, Brill S, van den Bos C, Eibl R: Investigations of fluid flow and cell proliferation of mesenchymal adipose-derived stem cells in small-scale, stirred, single-use bioreactors. Chem Ing Tech 2013, 85:95-102. doi:10.1186/1753-6561-7-S6-P2 Cite this article as: Schirmaier et al.: Expansion of mesenchymal adipose-tissue derived stem cells in a stirred single-use bioreactor under low-serum conditions. BMC Proceedings 2013 7(Suppl 6):P2. * Correspondence: *[email protected] 1 Zurich University of Applied Sciences, Institute of Biotechnology, Biochemical Engineering and Cell Cultivation Technique, 8820 Wädenswil, Switzerland Full list of author information is available at the end of the article Schirmaier et al. BMC Proceedings 2013, 7(Suppl 6):P2 http://www.biomedcentral.com/1753-6561/7/S6/P2 © 2013 Schirmaier et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Expansion of mesenchymal adipose-tissue derived stem cells in a stirred single-use bioreactor under low-serum conditions

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POSTER PRESENTATION Open Access

Expansion of mesenchymal adipose-tissuederived stem cells in a stirred single-usebioreactor under low-serum conditionsCarmen Schirmaier1*, Stephan C Kaiser1, Valentin Jossen1, Silke Brill2, Frank Jüngerkes2, Christian van den Bos2,Dieter Eibl1, Regine Eibl1

From 23rd European Society for Animal Cell Technology (ESACT) Meeting: Better Cells for Better HealthLille, France. 23-26 June 2013

BackgroundThe need for human mesenchymal stem cells (hMSCs)has increased enormously in recent years due to theirimportant therapeutic potential. Efficient cell expansionis essential to providing clinically relevant cell numbers.Such cell quantities can be manufactured by means ofscalable microcarrier (MC)-supported cultivations instirred single-use bioreactors.

Materials and methodsPreliminary tests in disposable-spinners (100 mL culturevolume, Corning) were used to determine two suitablemedia and MC-types for serum reduced expansions(< 10%) of human adipose tissue-derived stem cells(hADSCs; passage 2, Lonza). Using such optimizedmedia-MC-combinations, hADSCs expanded 30 to 40-fold, which compares well with expansion rates in planarculture. Based on computational fluid dynamics simula-tions and suspension analyses in spinners [1], optimaloperating parameters were determined in a BIOSTAT®

UniVessel® SU 2 L (2 L culture volume, Sartorius StedimBiotech).

ResultsIn subsequent batch tests with the BIOSTAT UniVessel®

SU 2 L, expansion rates of between 30 and 40-fold werereached and up to 4.4·108 cells with a cell viabilityexceeding 98% were harvested. Flow cytometry testsdemonstrated typical marker profiles following cell

expansion and harvest. A 40-fold expansion ratedelivered a total of 1·1010 cells in a first cultivationwith the BIOSTAT® CultiBag STR 50 L (35 L culturevolume, Sartorius Stedim Biotech).

ConclusionsIn summary, the foundations for successfully expandingtherapeutic stem cells in truly scalable systems havebeen laid. Strategies ensuring expansion rates between60 and 70-fold and, thus, generating cell amounts over1010 are now in preparation.

AcknowledgementsThis work is part of the project “Development of a technology platform for ascalable production of therapeutically relevant stem cells” (No. 12893.1VOUCH-LS). It is supported by the Commission for Technology andInnovation (CTI, Switzerland). The authors would like to thank the CTI forpartially financing the investigations presented.

Authors’ details1Zurich University of Applied Sciences, Institute of Biotechnology,Biochemical Engineering and Cell Cultivation Technique, 8820 Wädenswil,Switzerland. 2Lonza Cologne GmbH, 50829 Cologne, Germany.

Published: 4 December 2013

Reference1. Kaiser S C, Jossen V, Schirmaier C, Eibl D, Brill S, van den Bos C, Eibl R:

Investigations of fluid flow and cell proliferation of mesenchymaladipose-derived stem cells in small-scale, stirred, single-use bioreactors.Chem Ing Tech 2013, 85:95-102.

doi:10.1186/1753-6561-7-S6-P2Cite this article as: Schirmaier et al.: Expansion of mesenchymaladipose-tissue derived stem cells in a stirred single-use bioreactorunder low-serum conditions. BMC Proceedings 2013 7(Suppl 6):P2.

* Correspondence: *[email protected] University of Applied Sciences, Institute of Biotechnology,Biochemical Engineering and Cell Cultivation Technique, 8820 Wädenswil,SwitzerlandFull list of author information is available at the end of the article

Schirmaier et al. BMC Proceedings 2013, 7(Suppl 6):P2http://www.biomedcentral.com/1753-6561/7/S6/P2

© 2013 Schirmaier et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the CreativeCommons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, andreproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.