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Comparison of cultivation in Techne spinner, Bellco spinner, shake flask and T-flask of human embryonic stem cells
KTH, School of Biotechnology (BIO), Bioprocess Technology. (Cell Technology Group)ORCID iD: 0000-0002-5370-4621
KTH, School of Biotechnology (BIO), Bioprocess Technology. (Cell Technology Group)
KTH, School of Biotechnology (BIO), Bioprocess Technology. (Cell Technology Group)
Karolinska Institute, Department of Neurobiology, Care Sciences and Society. (Division of Alzheimer Neurobiology)
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2010 (English)In: Proceedings of the SBE's Second International Conference on Stem Cell Engineering, 2010Conference paper, Published paper (Other academic)
Abstract [en]

The recent progress in regenerative medicine indicates that pluripotent human embryonic stem cells (hESCs) may hold great potential providing cellular models for drug development and screening, modelling diseases as well as aid in the development of future cell-based therapies for neurodegenerative disorders, such as Alzheimer’s and Parkinson’s disease. Crucial to the success of generating specialized cell populations, is an understanding of the mechanisms, which influence the control of cell growth and differentiation by extrinsic and intrinsic factors. Nowadays, a limitation for the use of hESCs is the lack of proliferation methods in large scale. The purpose of the present work was to study several cultivation systems, which could potentially provide large-scale cultivation processes suitable for human therapy applications. Pluripotent human embryonic stem cells (hESCs), isolated from the inner cell mass of the blastocyst, were cultivated undifferentiated as embryoids bodies, i.e. large spherical aggregates of cells, in absence of serum and feeder layer. The cell growth and culture behavior in T-falsk, orbitally agitated shake flask, Bellco stirred spinner and Techne stirred spinner were observed. In Bellco spinner, the cells were agitated by a rotating impeller providing a movement comparable to stirred bioreactors. In Techne spinner, a slow and gentle orbital movement provided by a rotating bulb-ended stirrer maintained the cells in suspension. The design of this latter spinner allowed lower shear stress in comparison to Bellco spinner and shake flask. It was observed that the cell growth was fastest in Techne spinner followed by cultivation in T-flask and then cultivation in shake flask. Cultivating in Bellco spinner resulted in embryoid dissociation and viability decrease after 14 days. A larger number of single cells, i.e. cells not growing in aggregates, was observed in the static T-flask culture compared to the agitated systems, i.e. shake flask, Bellco spinner or Techne spinner. Probably the agitation promoted the spontaneous aggregation of the cells in spheres. In particular the Techne spinner allowed the most perfect spherical form among the different compared systems. Finally it was observed that hypoxia with 4 % oxygen concentration improved significantly the growth in Techne spinner or T-flask in comparison with normoxia with 21 % oxygen concentration. It was concluded that cultivation in Techne spinner under hypoxia was the most favorable condition among the ones studied here. The agitation provided by Techne spinner improved the cell growth in comparison with static system (T-flask). However using the other agitated systems, shake flask and Bellco spinner, was not comparably beneficial to the cell growth and viability, probably due to the higher shear stress of these systems compared to Techne spinner.

Place, publisher, year, edition, pages
2010.
Keyword [en]
human embryonic stem cell, bioreactor
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
SRA - Molecular Bioscience
Identifiers
URN: urn:nbn:se:kth:diva-87451OAI: oai:DiVA.org:kth-87451DiVA: diva2:501670
Conference
SBE's Second International Conference on Stem Cell Engineering, Boston, MA, USA, May 2-5, 2010
Note
QC 20120418Available from: 2012-02-14 Created: 2012-02-14 Last updated: 2012-04-18Bibliographically approved

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http://www.aiche.org/sbe/events/stemcelleng.aspx

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Chotteau, Veronique

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Chotteau, VeroniqueWang, JingjiaoTolf, ErikaWicklund, Linn
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