Expansion of 3D human induced pluripotent stem cell aggregates in bioreactors

Bioprocess intensification and scaling-up approaches

Tiago Aguiar, Rita Costa, Paula M. Alves, Anders Aspegren,, Patrícia Isabel Alves, Bernardo Abecassis, Émilie Arnault, Margarida Serra

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Human induced pluripotent stem cells (hiPSC) are attractive tools for drug screening and disease modeling and promising candidates for cell therapy applications. However, to achieve the high numbers of cells required for these purposes, scalable and clinical-grade technologies must be established. In this study, we use environmentally controlled stirred-tank bioreactors operating in perfusion as a powerful tool for bioprocess intensification of hiPSC production. We demonstrate the importance of controlling the dissolved oxygen concentration at low levels (4%) and perfusion at 1.3 day−1 dilution rate to improve hiPSC growth as aggregates in a xeno-free medium. This strategy allowed for increased cell specific growth rate, maximum volumetric concentrations (4.7 × 106 cell/mL) and expansion factors (approximately 19 in total cells), resulting in a 2.6-fold overall improvement in cell yields. Extensive cell characterization, including whole proteomic analysis, was performed to confirm that cells’ pluripotent phenotype was maintained during culture. A scalable protocol for continuous expansion of hiPSC aggregates in bioreactors was implemented using mechanical dissociation for aggregate disruption and cell passaging. A total expansion factor of 1100 in viable cells was obtained in 11 days of culture, while cells maintained their proliferation capacity, pluripotent phenotype and potential as well as genomic stability after 3 sequential passages in bioreactors.

Original languageEnglish
Pages (from-to)81-93
Number of pages13
JournalJournal of Biotechnology
Volume246
DOIs
Publication statusPublished - 20 Mar 2017

Fingerprint

Induced Pluripotent Stem Cells
Bioreactors
Stem cells
Cell culture
Cell growth
Dissolved oxygen
Dilution
Perfusion
Screening
Phenotype
Preclinical Drug Evaluations
Genomic Instability
Growth
Cell- and Tissue-Based Therapy
Proteomics
Pharmaceutical Preparations
Cell Culture Techniques
Cell Count
Oxygen
Technology

Keywords

  • Aggregate dissociation
  • Continuous expansion
  • Human induced pluripotent stem cells
  • Scale-up
  • Stirred tank bioreactors
  • Whole proteome

Cite this

@article{62244892881a4d36a2fbb2d3a5fdbfc9,
title = "Expansion of 3D human induced pluripotent stem cell aggregates in bioreactors: Bioprocess intensification and scaling-up approaches",
abstract = "Human induced pluripotent stem cells (hiPSC) are attractive tools for drug screening and disease modeling and promising candidates for cell therapy applications. However, to achieve the high numbers of cells required for these purposes, scalable and clinical-grade technologies must be established. In this study, we use environmentally controlled stirred-tank bioreactors operating in perfusion as a powerful tool for bioprocess intensification of hiPSC production. We demonstrate the importance of controlling the dissolved oxygen concentration at low levels (4{\%}) and perfusion at 1.3 day−1 dilution rate to improve hiPSC growth as aggregates in a xeno-free medium. This strategy allowed for increased cell specific growth rate, maximum volumetric concentrations (4.7 × 106 cell/mL) and expansion factors (approximately 19 in total cells), resulting in a 2.6-fold overall improvement in cell yields. Extensive cell characterization, including whole proteomic analysis, was performed to confirm that cells’ pluripotent phenotype was maintained during culture. A scalable protocol for continuous expansion of hiPSC aggregates in bioreactors was implemented using mechanical dissociation for aggregate disruption and cell passaging. A total expansion factor of 1100 in viable cells was obtained in 11 days of culture, while cells maintained their proliferation capacity, pluripotent phenotype and potential as well as genomic stability after 3 sequential passages in bioreactors.",
keywords = "Aggregate dissociation, Continuous expansion, Human induced pluripotent stem cells, Scale-up, Stirred tank bioreactors, Whole proteome",
author = "Tiago Aguiar and Rita Costa and Alves, {Paula M.} and Anders Aspegren, and Alves, {Patr{\'i}cia Isabel} and Bernardo Abecassis and {\'E}milie Arnault and Margarida Serra",
year = "2017",
month = "3",
day = "20",
doi = "10.1016/j.jbiotec.2017.01.004",
language = "English",
volume = "246",
pages = "81--93",
journal = "Journal of Biotechnology",
issn = "0168-1656",
publisher = "Elsevier Science B.V., Inc",

}

Expansion of 3D human induced pluripotent stem cell aggregates in bioreactors : Bioprocess intensification and scaling-up approaches. / Aguiar, Tiago; Costa, Rita; Alves, Paula M.; Aspegren, Anders; Alves, Patrícia Isabel; Abecassis, Bernardo; Arnault, Émilie; Serra, Margarida.

In: Journal of Biotechnology, Vol. 246, 20.03.2017, p. 81-93.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Expansion of 3D human induced pluripotent stem cell aggregates in bioreactors

T2 - Bioprocess intensification and scaling-up approaches

AU - Aguiar, Tiago

AU - Costa, Rita

AU - Alves, Paula M.

AU - Aspegren,, Anders

AU - Alves, Patrícia Isabel

AU - Abecassis, Bernardo

AU - Arnault, Émilie

AU - Serra, Margarida

PY - 2017/3/20

Y1 - 2017/3/20

N2 - Human induced pluripotent stem cells (hiPSC) are attractive tools for drug screening and disease modeling and promising candidates for cell therapy applications. However, to achieve the high numbers of cells required for these purposes, scalable and clinical-grade technologies must be established. In this study, we use environmentally controlled stirred-tank bioreactors operating in perfusion as a powerful tool for bioprocess intensification of hiPSC production. We demonstrate the importance of controlling the dissolved oxygen concentration at low levels (4%) and perfusion at 1.3 day−1 dilution rate to improve hiPSC growth as aggregates in a xeno-free medium. This strategy allowed for increased cell specific growth rate, maximum volumetric concentrations (4.7 × 106 cell/mL) and expansion factors (approximately 19 in total cells), resulting in a 2.6-fold overall improvement in cell yields. Extensive cell characterization, including whole proteomic analysis, was performed to confirm that cells’ pluripotent phenotype was maintained during culture. A scalable protocol for continuous expansion of hiPSC aggregates in bioreactors was implemented using mechanical dissociation for aggregate disruption and cell passaging. A total expansion factor of 1100 in viable cells was obtained in 11 days of culture, while cells maintained their proliferation capacity, pluripotent phenotype and potential as well as genomic stability after 3 sequential passages in bioreactors.

AB - Human induced pluripotent stem cells (hiPSC) are attractive tools for drug screening and disease modeling and promising candidates for cell therapy applications. However, to achieve the high numbers of cells required for these purposes, scalable and clinical-grade technologies must be established. In this study, we use environmentally controlled stirred-tank bioreactors operating in perfusion as a powerful tool for bioprocess intensification of hiPSC production. We demonstrate the importance of controlling the dissolved oxygen concentration at low levels (4%) and perfusion at 1.3 day−1 dilution rate to improve hiPSC growth as aggregates in a xeno-free medium. This strategy allowed for increased cell specific growth rate, maximum volumetric concentrations (4.7 × 106 cell/mL) and expansion factors (approximately 19 in total cells), resulting in a 2.6-fold overall improvement in cell yields. Extensive cell characterization, including whole proteomic analysis, was performed to confirm that cells’ pluripotent phenotype was maintained during culture. A scalable protocol for continuous expansion of hiPSC aggregates in bioreactors was implemented using mechanical dissociation for aggregate disruption and cell passaging. A total expansion factor of 1100 in viable cells was obtained in 11 days of culture, while cells maintained their proliferation capacity, pluripotent phenotype and potential as well as genomic stability after 3 sequential passages in bioreactors.

KW - Aggregate dissociation

KW - Continuous expansion

KW - Human induced pluripotent stem cells

KW - Scale-up

KW - Stirred tank bioreactors

KW - Whole proteome

UR - http://www.scopus.com/inward/record.url?scp=85013073444&partnerID=8YFLogxK

U2 - 10.1016/j.jbiotec.2017.01.004

DO - 10.1016/j.jbiotec.2017.01.004

M3 - Article

VL - 246

SP - 81

EP - 93

JO - Journal of Biotechnology

JF - Journal of Biotechnology

SN - 0168-1656

ER -