Finding the design space of a filtration-based operation for the concentration of human pluripotent stem cells

Bárbara Cunha, Ricardo J.S. Silva, Cláudia Correia, Alexey Koshkin, Paula M. Alves, Margarida Serra, Cristina Peixoto, Manuel J.T. Carrondo

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Process knowledge for designing robust and reproducible unit operations is essential, especially for complex biological systems. This work describes a shortcut approach for the design of tangential flow filtration for the concentration of human induced pluripotent stem cells (hiPSC), supported by design of experiments. Critical process parameters (CPP) of shear rate, permeate flux and cell load were considered, and their impact on hiPSC recovery yield and viability was studied. A full factorial design confirmed significant interaction effects between all CPP, affecting both responses. The developed statistical model predicted that high shear rate (3000 s−1), permeate flux (250 LMH) and medium cell load (2 × 106 cell/cm2) would maximize both cell recovery yield and viability, where over 80% of hiPSC were recovered after a volume reduction factor of 20 with high viability (over 93%). Such conditions were validated experimentally, and by performing a robustness analysis, the success rate of these operating conditions was assessed (65–70%). A parametric study was then conducted, identifying that increasing the shear rate (up to 3370 s−1) allowed to achieve the specified requirements for cell recovery yield (>80%) and viability (>90%) in 100% of the cases and no impact in hiPSC's identity, proliferation capacity and pluripotency was observed.

Original languageEnglish
Pages (from-to)399-407
Number of pages9
JournalJournal of Membrane Science
Volume542
DOIs
Publication statusPublished - 1 Jan 2017

Fingerprint

Pluripotent Stem Cells
stem cells
Induced Pluripotent Stem Cells
Stem cells
Shear deformation
viability
Recovery
cells
Fluxes
recovery
shear
Biological systems
Design of experiments
factorial design
experiment design
Statistical Models
flux (rate)
requirements
interactions

Keywords

  • Design of experiments
  • Downstream processing
  • Human induced pluripotent stem cells
  • Quality by design
  • Tangential flow filtration

Cite this

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title = "Finding the design space of a filtration-based operation for the concentration of human pluripotent stem cells",
abstract = "Process knowledge for designing robust and reproducible unit operations is essential, especially for complex biological systems. This work describes a shortcut approach for the design of tangential flow filtration for the concentration of human induced pluripotent stem cells (hiPSC), supported by design of experiments. Critical process parameters (CPP) of shear rate, permeate flux and cell load were considered, and their impact on hiPSC recovery yield and viability was studied. A full factorial design confirmed significant interaction effects between all CPP, affecting both responses. The developed statistical model predicted that high shear rate (3000 s−1), permeate flux (250 LMH) and medium cell load (2 × 106 cell/cm2) would maximize both cell recovery yield and viability, where over 80{\%} of hiPSC were recovered after a volume reduction factor of 20 with high viability (over 93{\%}). Such conditions were validated experimentally, and by performing a robustness analysis, the success rate of these operating conditions was assessed (65–70{\%}). A parametric study was then conducted, identifying that increasing the shear rate (up to 3370 s−1) allowed to achieve the specified requirements for cell recovery yield (>80{\%}) and viability (>90{\%}) in 100{\%} of the cases and no impact in hiPSC's identity, proliferation capacity and pluripotency was observed.",
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Finding the design space of a filtration-based operation for the concentration of human pluripotent stem cells. / Cunha, Bárbara; Silva, Ricardo J.S.; Correia, Cláudia; Koshkin, Alexey; Alves, Paula M.; Serra, Margarida; Peixoto, Cristina; Carrondo, Manuel J.T.

In: Journal of Membrane Science, Vol. 542, 01.01.2017, p. 399-407.

Research output: Contribution to journalArticle

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T1 - Finding the design space of a filtration-based operation for the concentration of human pluripotent stem cells

AU - Cunha, Bárbara

AU - Silva, Ricardo J.S.

AU - Correia, Cláudia

AU - Koshkin, Alexey

AU - Alves, Paula M.

AU - Serra, Margarida

AU - Peixoto, Cristina

AU - Carrondo, Manuel J.T.

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