Details
| Original language | English |
|---|---|
| Pages (from-to) | 1289-1301 |
| Number of pages | 13 |
| Journal | Stem cells translational medicine |
| Volume | 5 |
| Issue number | 10 |
| Publication status | Published - 1 Jul 2016 |
Abstract
Theroutine applicationofhumanpluripotent stemcells (hPSCs)andtheirderivatives inbiomedicineanddrug discoverywill require the constant supply of high-quality cells by defined processes. Culturing hPSCs as cellonly aggregates in (three-dimensional [3D]) suspension has the potential to overcome numerous limitations of conventional surface-adherent (two-dimensional [2D]) cultivation. Utilizing single-use instrumented stirred-tank bioreactors, we showed that perfusion resulted in a more homogeneous culture environment and enabled superior cell densities of 2.85×106 cells permilliliter and 47% higher cell yields compared with conventional repeated batch cultures. Flow cytometry, quantitative reverse-transcriptase polymerase chain reaction, and global gene expression analysis revealed a high similarity across 3D suspension and 2D precultures, underscoring that matrix-free hPSC culture efficiently supportsmaintenance of pluripotency. Interestingly, physiological data and gene expression assessment indicated distinct changes of the cells’ energy metabolism, suggesting a culture-induced switch from glycolysis to oxidative phosphorylation in the absence of hPSC differentiation. Our data highlight the plasticity of hPSCs’ energy metabolism and provide clear physiological and molecular targets for process monitoring and further development. This study paves the way toward more efficientGMP-compliant cell production and underscores the enormous process development potential of hPSCs in suspension culture.
Keywords
- Human pluripotent stem cell expansion, Induced pluripotent stem cells, Metabolism, Perfusion, Single-use stirred tank bioreactors, Three-dimensional suspension culture
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Developmental Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
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In: Stem cells translational medicine, Vol. 5, No. 10, 01.07.2016, p. 1289-1301.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Impact of Feeding Strategies on the Scalable Expansion of Human Pluripotent Stem Cells in Single‐Use Stirred Tank Bioreactors
AU - Kropp, Christina
AU - Kempf, Henning
AU - Halloin, Caroline
AU - Robles-Diaz, Diana
AU - Franke, Annika
AU - Scheper, Thomas
AU - Kinast, Katharina
AU - Knorpp, Thomas
AU - Joos, Thomas O.
AU - Haverich, Axel
AU - Martin, Ulrich
AU - Zweigerdt, Robert
AU - Olmera, Ruth
N1 - © 2016 AlphaMed Press
PY - 2016/7/1
Y1 - 2016/7/1
N2 - Theroutine applicationofhumanpluripotent stemcells (hPSCs)andtheirderivatives inbiomedicineanddrug discoverywill require the constant supply of high-quality cells by defined processes. Culturing hPSCs as cellonly aggregates in (three-dimensional [3D]) suspension has the potential to overcome numerous limitations of conventional surface-adherent (two-dimensional [2D]) cultivation. Utilizing single-use instrumented stirred-tank bioreactors, we showed that perfusion resulted in a more homogeneous culture environment and enabled superior cell densities of 2.85×106 cells permilliliter and 47% higher cell yields compared with conventional repeated batch cultures. Flow cytometry, quantitative reverse-transcriptase polymerase chain reaction, and global gene expression analysis revealed a high similarity across 3D suspension and 2D precultures, underscoring that matrix-free hPSC culture efficiently supportsmaintenance of pluripotency. Interestingly, physiological data and gene expression assessment indicated distinct changes of the cells’ energy metabolism, suggesting a culture-induced switch from glycolysis to oxidative phosphorylation in the absence of hPSC differentiation. Our data highlight the plasticity of hPSCs’ energy metabolism and provide clear physiological and molecular targets for process monitoring and further development. This study paves the way toward more efficientGMP-compliant cell production and underscores the enormous process development potential of hPSCs in suspension culture.
AB - Theroutine applicationofhumanpluripotent stemcells (hPSCs)andtheirderivatives inbiomedicineanddrug discoverywill require the constant supply of high-quality cells by defined processes. Culturing hPSCs as cellonly aggregates in (three-dimensional [3D]) suspension has the potential to overcome numerous limitations of conventional surface-adherent (two-dimensional [2D]) cultivation. Utilizing single-use instrumented stirred-tank bioreactors, we showed that perfusion resulted in a more homogeneous culture environment and enabled superior cell densities of 2.85×106 cells permilliliter and 47% higher cell yields compared with conventional repeated batch cultures. Flow cytometry, quantitative reverse-transcriptase polymerase chain reaction, and global gene expression analysis revealed a high similarity across 3D suspension and 2D precultures, underscoring that matrix-free hPSC culture efficiently supportsmaintenance of pluripotency. Interestingly, physiological data and gene expression assessment indicated distinct changes of the cells’ energy metabolism, suggesting a culture-induced switch from glycolysis to oxidative phosphorylation in the absence of hPSC differentiation. Our data highlight the plasticity of hPSCs’ energy metabolism and provide clear physiological and molecular targets for process monitoring and further development. This study paves the way toward more efficientGMP-compliant cell production and underscores the enormous process development potential of hPSCs in suspension culture.
KW - Human pluripotent stem cell expansion
KW - Induced pluripotent stem cells
KW - Metabolism
KW - Perfusion
KW - Single-use stirred tank bioreactors
KW - Three-dimensional suspension culture
UR - http://www.scopus.com/inward/record.url?scp=84988596815&partnerID=8YFLogxK
U2 - 10.5966/sctm.2015-0253
DO - 10.5966/sctm.2015-0253
M3 - Article
C2 - 27369897
AN - SCOPUS:84988596815
VL - 5
SP - 1289
EP - 1301
JO - Stem cells translational medicine
JF - Stem cells translational medicine
SN - 2157-6564
IS - 10
ER -