Filtration methodologies for the clarification and concentration of human mesenchymal stem cells

Bárbara Cunha, Cristina Peixoto, Marta M. Silva, Manuel José Teixeira Carrondo, Margarida Serra, Paula M. Alves

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Currently human mesenchymal stem cells (hMSC) are expanded using microcarrier-based stirred culture systems from one to hundreds of liters of culture volume to guarantee the required cell numbers to be delivered to the clinic. Such culture volumes need to be clarified, ensuring efficient removal of microcarriers, and concentrated without compromising the cells' characteristics. The aim of this work was to evaluate the applicability of filtration methodologies, as dead end filtration and tangential flow filtration, for the clarification and concentration of hMSC, respectively.Different process variables and their impact on hMSC quality were evaluated, showing that polypropylene filters with pore sizes higher than 75μm can ensure the removal of microcarriers from the cell suspension bulk, without compromising cells' recovery or viability. Furthermore, hMSC could be successfully concentrated up to a factor of ten while maintaining their identity, potency and high cell viability, allowing for the recovery of over 80% of viable cells; an initial cell concentration higher than 2×105 cell/mL, and polysulfone membranes with pore sizes higher than 0.45μm were identified to be key conditions to obtain such concentration factors; shear rate and permeate flux were also shown to impact the cells' recovery yields, viability and quality.

Original languageEnglish
Pages (from-to)117-129
Number of pages13
JournalJournal of Membrane Science
Volume478
DOIs
Publication statusPublished - 5 Mar 2015

Fingerprint

stem cells
Stem cells
Mesenchymal Stromal Cells
methodology
cells
Recovery
Pore size
viability
Polysulfones
Polypropylenes
recovery
Shear deformation
Suspensions
Cells
Fluxes
Membranes
porosity
polypropylene
Cell Survival
Cell Count

Keywords

  • Cell concentration
  • Downstream processing
  • Human mesenchymal stem cells
  • Microcarriers
  • Tangential flow filtration

Cite this

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abstract = "Currently human mesenchymal stem cells (hMSC) are expanded using microcarrier-based stirred culture systems from one to hundreds of liters of culture volume to guarantee the required cell numbers to be delivered to the clinic. Such culture volumes need to be clarified, ensuring efficient removal of microcarriers, and concentrated without compromising the cells' characteristics. The aim of this work was to evaluate the applicability of filtration methodologies, as dead end filtration and tangential flow filtration, for the clarification and concentration of hMSC, respectively.Different process variables and their impact on hMSC quality were evaluated, showing that polypropylene filters with pore sizes higher than 75μm can ensure the removal of microcarriers from the cell suspension bulk, without compromising cells' recovery or viability. Furthermore, hMSC could be successfully concentrated up to a factor of ten while maintaining their identity, potency and high cell viability, allowing for the recovery of over 80{\%} of viable cells; an initial cell concentration higher than 2×105 cell/mL, and polysulfone membranes with pore sizes higher than 0.45μm were identified to be key conditions to obtain such concentration factors; shear rate and permeate flux were also shown to impact the cells' recovery yields, viability and quality.",
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Filtration methodologies for the clarification and concentration of human mesenchymal stem cells. / Cunha, Bárbara; Peixoto, Cristina; Silva, Marta M.; Carrondo, Manuel José Teixeira; Serra, Margarida; Alves, Paula M.

In: Journal of Membrane Science, Vol. 478, 05.03.2015, p. 117-129.

Research output: Contribution to journalArticle

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