Growth Behavior of Human Adipose Tissue-Derived Stromal/Stem Cells at Small Scale: Numerical and Experimental Investigations
Human adipose tissue-derived stromal/stem cells (hASCs) are a valuable source of cells for clinical applications, especially in the field of regenerative medicine. Therefore, it comes as no surprise that the interest in hASCs has greatly increased over the last decade. However, in order to use hASCs...
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MDPI AG
2018-12-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/5/4/106 |
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| author | Valentin Jossen Regine Eibl Matthias Kraume Dieter Eibl |
| author_facet | Valentin Jossen Regine Eibl Matthias Kraume Dieter Eibl |
| author_sort | Valentin Jossen |
| collection | DOAJ |
| description | Human adipose tissue-derived stromal/stem cells (hASCs) are a valuable source of cells for clinical applications, especially in the field of regenerative medicine. Therefore, it comes as no surprise that the interest in hASCs has greatly increased over the last decade. However, in order to use hASCs in clinically relevant numbers, in vitro expansion is required. Single-use stirred bioreactors in combination with microcarriers (MCs) have shown themselves to be suitable systems for this task. However, hASCs tend to be less robust, and thus, more shear sensitive than conventional production cell lines for therapeutic antibodies and vaccines (e.g., Chinese Hamster Ovary cells CHO, Baby Hamster Kidney cells BHK), for which these bioreactors were originally designed. Hence, the goal of this study was to investigate the influence of different shear stress levels on the growth of humane telomerase reversed transcriptase immortalized hASCs (hTERT-ASC) and aggregate formation in stirred single-use systems at the mL scale: the 125 mL (=SP100) and the 500 mL (=SP300) disposable Corning<sup>®</sup> spinner flask. Computational fluid dynamics (CFD) simulations based on an Euler⁻Euler and Euler⁻Lagrange approach were performed to predict the hydrodynamic stresses (0.06⁻0.87 Pa), the residence times (0.4⁻7.3 s), and the circulation times (1.6⁻16.6 s) of the MCs in different shear zones for different impeller speeds and the suspension criteria (<i>Ns1u, Ns1</i>). The numerical findings were linked to experimental data from cultivations studies to develop, for the first time, an unstructured, segregated mathematical growth model for hTERT-ASCs. While the 125 mL spinner flask with 100 mL working volume (SP100) provided up to 1.68 × 10<sup>5</sup> hTERT-ASC/cm<sup>2</sup> (=0.63 × 10<sup>6</sup> living hTERT-ASCs/mL, EF 56) within eight days, the peak living cell density of the 500 mL spinner flask with 300 mL working volume (SP300) was 2.46 × 10<sup>5</sup> hTERT-ASC/cm<sup>2</sup> (=0.88 × 10<sup>6</sup> hTERT-ASCs/mL, EF 81) and was achieved on day eight. Optimal cultivation conditions were found for <i>Ns1u</i> < <i>N</i> < <i>Ns1</i>, which corresponded to specific power inputs of 0.3⁻1.1 W/m<sup>3</sup>. The established growth model delivered reliable predictions for cell growth on the MCs with an accuracy of 76⁻96% for both investigated spinner flask types. |
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| spelling | doaj.art-711583f7577543c588fd01b697f010352023-09-02T13:21:57ZengMDPI AGBioengineering2306-53542018-12-015410610.3390/bioengineering5040106bioengineering5040106Growth Behavior of Human Adipose Tissue-Derived Stromal/Stem Cells at Small Scale: Numerical and Experimental InvestigationsValentin Jossen0Regine Eibl1Matthias Kraume2Dieter Eibl3Institute of Chemistry and Biotechnology, Zurich University of Applied Sciences, 8820 Wädenswil, SwitzerlandInstitute of Chemistry and Biotechnology, Zurich University of Applied Sciences, 8820 Wädenswil, SwitzerlandDepartment of Process Engineering, Technical University of Berlin, 10623 Berlin, Germany, <email>matthias.kraume@tu-berlin.de</email>Institute of Chemistry and Biotechnology, Zurich University of Applied Sciences, 8820 Wädenswil, SwitzerlandHuman adipose tissue-derived stromal/stem cells (hASCs) are a valuable source of cells for clinical applications, especially in the field of regenerative medicine. Therefore, it comes as no surprise that the interest in hASCs has greatly increased over the last decade. However, in order to use hASCs in clinically relevant numbers, in vitro expansion is required. Single-use stirred bioreactors in combination with microcarriers (MCs) have shown themselves to be suitable systems for this task. However, hASCs tend to be less robust, and thus, more shear sensitive than conventional production cell lines for therapeutic antibodies and vaccines (e.g., Chinese Hamster Ovary cells CHO, Baby Hamster Kidney cells BHK), for which these bioreactors were originally designed. Hence, the goal of this study was to investigate the influence of different shear stress levels on the growth of humane telomerase reversed transcriptase immortalized hASCs (hTERT-ASC) and aggregate formation in stirred single-use systems at the mL scale: the 125 mL (=SP100) and the 500 mL (=SP300) disposable Corning<sup>®</sup> spinner flask. Computational fluid dynamics (CFD) simulations based on an Euler⁻Euler and Euler⁻Lagrange approach were performed to predict the hydrodynamic stresses (0.06⁻0.87 Pa), the residence times (0.4⁻7.3 s), and the circulation times (1.6⁻16.6 s) of the MCs in different shear zones for different impeller speeds and the suspension criteria (<i>Ns1u, Ns1</i>). The numerical findings were linked to experimental data from cultivations studies to develop, for the first time, an unstructured, segregated mathematical growth model for hTERT-ASCs. While the 125 mL spinner flask with 100 mL working volume (SP100) provided up to 1.68 × 10<sup>5</sup> hTERT-ASC/cm<sup>2</sup> (=0.63 × 10<sup>6</sup> living hTERT-ASCs/mL, EF 56) within eight days, the peak living cell density of the 500 mL spinner flask with 300 mL working volume (SP300) was 2.46 × 10<sup>5</sup> hTERT-ASC/cm<sup>2</sup> (=0.88 × 10<sup>6</sup> hTERT-ASCs/mL, EF 81) and was achieved on day eight. Optimal cultivation conditions were found for <i>Ns1u</i> < <i>N</i> < <i>Ns1</i>, which corresponded to specific power inputs of 0.3⁻1.1 W/m<sup>3</sup>. The established growth model delivered reliable predictions for cell growth on the MCs with an accuracy of 76⁻96% for both investigated spinner flask types.https://www.mdpi.com/2306-5354/5/4/106computational fluid dynamicshuman adipose tissue-derived stromal/stem cellshumane telomerase reversed transcriptase immortalized hASCsmicrocarriersegregated growth modelEuler–Euler and Euler–Lagrange approachesparticle image velocimetry/shadowgraphy measurements |
| spellingShingle | Valentin Jossen Regine Eibl Matthias Kraume Dieter Eibl Growth Behavior of Human Adipose Tissue-Derived Stromal/Stem Cells at Small Scale: Numerical and Experimental Investigations Bioengineering computational fluid dynamics human adipose tissue-derived stromal/stem cells humane telomerase reversed transcriptase immortalized hASCs microcarrier segregated growth model Euler–Euler and Euler–Lagrange approaches particle image velocimetry/shadowgraphy measurements |
| title | Growth Behavior of Human Adipose Tissue-Derived Stromal/Stem Cells at Small Scale: Numerical and Experimental Investigations |
| title_full | Growth Behavior of Human Adipose Tissue-Derived Stromal/Stem Cells at Small Scale: Numerical and Experimental Investigations |
| title_fullStr | Growth Behavior of Human Adipose Tissue-Derived Stromal/Stem Cells at Small Scale: Numerical and Experimental Investigations |
| title_full_unstemmed | Growth Behavior of Human Adipose Tissue-Derived Stromal/Stem Cells at Small Scale: Numerical and Experimental Investigations |
| title_short | Growth Behavior of Human Adipose Tissue-Derived Stromal/Stem Cells at Small Scale: Numerical and Experimental Investigations |
| title_sort | growth behavior of human adipose tissue derived stromal stem cells at small scale numerical and experimental investigations |
| topic | computational fluid dynamics human adipose tissue-derived stromal/stem cells humane telomerase reversed transcriptase immortalized hASCs microcarrier segregated growth model Euler–Euler and Euler–Lagrange approaches particle image velocimetry/shadowgraphy measurements |
| url | https://www.mdpi.com/2306-5354/5/4/106 |
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