Development of a device useful to reproducibly produce large quantities of viable and uniform stem cell spheroids with controlled diameters

Monize Caiado Decarli, Mateus Vidigal de Castro, Júlia Adami Nogueira, Mariana Harue T Nagahara, Cecília Buzatto Westin, Alexandre Leite R de Oliveira, Jorge Vicente L da Silva, Lorenzo Moroni, Carlos Mota, Ângela Maria Moraes*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Three-dimensional cellular aggregates can mimic the natural microenvironment of tissues and organs and obtaining them through controlled and reproducible processes is mandatory for scaling up and implementing drug cytotoxicity and efficacy tests, as well as tissue engineering protocols. The purpose of this work was to develop and evaluate the performance of a device with two different geometries fabricated by additive manufacturing. The methodology was based on casting a microwell array insert using a non-adhesive hydrogel to obtain highly regular microcavities to standardize spheroid formation and morphology. Spheroids of dental pulp stem cells, bone marrow stromal cells and embryonic stem cells showing high cell viability and average diameters of around 253, 220, and 500 μm, respectively, were produced using the device with the geometry considered most adequate. The cell aggregates showed sphericity indexes above 0.9 and regular surfaces (solidity index higher than 0.96). Around 1000 spheroids could be produced in a standard six-well plate. Overall, these results show that this method facilitates obtaining a large number of uniform, viable spheroids with pre-specified average diameters and through a low-cost and reproducible process for a myriad of applications.

Original languageEnglish
Article number112685
Number of pages14
JournalBiomaterials Advances
Volume135
Early online date31 Jan 2022
DOIs
Publication statusPublished - Apr 2022

Keywords

  • 3D cell culture
  • 3D cell model
  • CULTURE
  • Cell spheroid
  • DESIGN
  • DRUG
  • In vitro model
  • MODELS
  • MULTICELLULAR SPHEROIDS
  • Microwell array
  • NECROSIS
  • ON-A-CHIP
  • Stem cells
  • additive manufacturing

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