Cell spheroids as a versatile research platform: formation mechanisms, high throughput production, characterization and applications

Three-dimensional (3D) cell culture has tremendous advantages to closely mimic the architecture and microenvironment of healthy tissue and organs, as well as of solid tumors. Spheroids are currently the most attractive 3D model to produce uniform reproducible cell structures as well as a potential b...

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Published inBiofabrication Vol. 13; no. 3; p. 32002
Main Authors Decarli, Monize Caiado, Amaral, Robson, Santos, Diogo Peres Dos, Tofani, Larissa Bueno, Katayama, Eric, Rezende, Rodrigo Alvarenga, Silva, Jorge Vicente Lopes da, Swiech, Kamilla, Suazo, Claudio Alberto Torres, Mota, Carlos, Moroni, Lorenzo, Moraes, Ângela Maria
Format Journal Article
LanguageEnglish
Published England 01.07.2021
Subjects
Bioprinting
Cell Culture Techniques
Hydrogels
Spheroids, Cellular
Tissue Engineering
tissue engineering
drug screening
3D cell culture
3D bioprinting
cell aggregation
cell spheroids
spheroids as in vitro models
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Abstract Three-dimensional (3D) cell culture has tremendous advantages to closely mimic the architecture and microenvironment of healthy tissue and organs, as well as of solid tumors. Spheroids are currently the most attractive 3D model to produce uniform reproducible cell structures as well as a potential basis for engineering large tissues and complex organs. In this review we discuss, from an engineering perspective, processes to obtain uniform 3D cell spheroids, comparing dynamic and static cultures and considering aspects such as mass transfer and shear stress. In addition, computational and mathematical modeling of complex cell spheroid systems are discussed. The non-cell-adhesive hydrogel-based method and dynamic cell culture in bioreactors are focused in detail and the myriad of developed spheroid characterization techniques is presented. The main bottlenecks and weaknesses are discussed, especially regarding the analysis of morphological parameters, cell quantification and viability, gene expression profiles, metabolic behavior and high-content analysis. Finally, a vast set of applications of spheroids as tools for study model systems is examined, including drug screening, tissue formation, pathologies development, tissue engineering and biofabrication, 3D bioprinting and microfluidics, together with their use in high-throughput platforms.
AbstractList Abstract Three-dimensional (3D) cell culture has tremendous advantages to closely mimic the in vivo architecture and microenvironment of healthy tissue and organs, as well as of solid tumors. Spheroids are currently the most attractive 3D model to produce uniform reproducible cell structures as well as a potential basis for engineering large tissues and complex organs. In this review we discuss, from an engineering perspective, processes to obtain uniform 3D cell spheroids, comparing dynamic and static cultures and considering aspects such as mass transfer and shear stress. In addition, computational and mathematical modeling of complex cell spheroid systems are discussed. The non-cell-adhesive hydrogel-based method and dynamic cell culture in bioreactors are focused in detail and the myriad of developed spheroid characterization techniques is presented. The main bottlenecks and weaknesses are discussed, especially regarding the analysis of morphological parameters, cell quantification and viability, gene expression profiles, metabolic behavior and high-content analysis. Finally, a vast set of applications of spheroids as tools for in vitro study model systems is examined, including drug screening, tissue formation, pathologies development, tissue engineering and biofabrication, 3D bioprinting and microfluidics, together with their use in high-throughput platforms.
Three-dimensional (3D) cell culture has tremendous advantages to closely mimic the architecture and microenvironment of healthy tissue and organs, as well as of solid tumors. Spheroids are currently the most attractive 3D model to produce uniform reproducible cell structures as well as a potential basis for engineering large tissues and complex organs. In this review we discuss, from an engineering perspective, processes to obtain uniform 3D cell spheroids, comparing dynamic and static cultures and considering aspects such as mass transfer and shear stress. In addition, computational and mathematical modeling of complex cell spheroid systems are discussed. The non-cell-adhesive hydrogel-based method and dynamic cell culture in bioreactors are focused in detail and the myriad of developed spheroid characterization techniques is presented. The main bottlenecks and weaknesses are discussed, especially regarding the analysis of morphological parameters, cell quantification and viability, gene expression profiles, metabolic behavior and high-content analysis. Finally, a vast set of applications of spheroids as tools for study model systems is examined, including drug screening, tissue formation, pathologies development, tissue engineering and biofabrication, 3D bioprinting and microfluidics, together with their use in high-throughput platforms.
Author Moroni, Lorenzo
Mota, Carlos
Decarli, Monize Caiado
Moraes, Ângela Maria
Katayama, Eric
Rezende, Rodrigo Alvarenga
Tofani, Larissa Bueno
Swiech, Kamilla
Santos, Diogo Peres Dos
Amaral, Robson
Silva, Jorge Vicente Lopes da
Suazo, Claudio Alberto Torres
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  orcidid: 0000-0002-7100-3394
  surname: Decarli
  fullname: Decarli, Monize Caiado
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– sequence: 2
  givenname: Robson
  surname: Amaral
  fullname: Amaral, Robson
  organization: Department of Pharmaceutical Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Avenida do Café, no number, 14040-903 Ribeirão Preto, SP, Brazil
– sequence: 3
  givenname: Diogo Peres Dos
  surname: Santos
  fullname: Santos, Diogo Peres Dos
  organization: Departament of Chemical Engineering, Federal University of São Carlos, Rod. Washington Luiz (SP-310), km 235, 13565-905 São Carlos, SP, Brazil
– sequence: 4
  givenname: Larissa Bueno
  surname: Tofani
  fullname: Tofani, Larissa Bueno
  organization: Department of Pharmaceutical Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Avenida do Café, no number, 14040-903 Ribeirão Preto, SP, Brazil
– sequence: 5
  givenname: Eric
  surname: Katayama
  fullname: Katayama, Eric
  organization: Departament of Chemical Engineering, Federal University of São Carlos, Rod. Washington Luiz (SP-310), km 235, 13565-905 São Carlos, SP, Brazil
– sequence: 6
  givenname: Rodrigo Alvarenga
  surname: Rezende
  fullname: Rezende, Rodrigo Alvarenga
  organization: Renato Archer Information Technology Center CTI, Rod. Dom Pedro I (SP-65), km 143,6, Amarais13069-901 Campinas, SP, Brazil
– sequence: 7
  givenname: Jorge Vicente Lopes da
  surname: Silva
  fullname: Silva, Jorge Vicente Lopes da
  organization: Renato Archer Information Technology Center CTI, Rod. Dom Pedro I (SP-65), km 143,6, Amarais13069-901 Campinas, SP, Brazil
– sequence: 8
  givenname: Kamilla
  surname: Swiech
  fullname: Swiech, Kamilla
  organization: Department of Pharmaceutical Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Avenida do Café, no number, 14040-903 Ribeirão Preto, SP, Brazil
– sequence: 9
  givenname: Claudio Alberto Torres
  surname: Suazo
  fullname: Suazo, Claudio Alberto Torres
  organization: Departament of Chemical Engineering, Federal University of São Carlos, Rod. Washington Luiz (SP-310), km 235, 13565-905 São Carlos, SP, Brazil
– sequence: 10
  givenname: Carlos
  surname: Mota
  fullname: Mota, Carlos
  organization: MERLN Institute for Technology-Inspired Regenerative Medicine, Department of Complex Tissue Regeneration, Maastricht University, Universiteitssingel, 40, 6229 ER Maastricht, The Netherlands
– sequence: 11
  givenname: Lorenzo
  orcidid: 0000-0003-1298-6025
  surname: Moroni
  fullname: Moroni, Lorenzo
  organization: MERLN Institute for Technology-Inspired Regenerative Medicine, Department of Complex Tissue Regeneration, Maastricht University, Universiteitssingel, 40, 6229 ER Maastricht, The Netherlands
– sequence: 12
  givenname: Ângela Maria
  orcidid: 0000-0002-5813-332X
  surname: Moraes
  fullname: Moraes, Ângela Maria
  organization: Department of Engineering of Materials and of Bioprocesses, School of Chemical Engineering, University of Campinas, Av. Albert Einstein, 500, 13083-852 Campinas, SP, Brazil
BackLink https://www.ncbi.nlm.nih.gov/pubmed/33592595$$D View this record in MEDLINE/PubMed
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Issue 3
Keywords tissue engineering
drug screening
3D cell culture
3D bioprinting
cell aggregation
cell spheroids
spheroids as in vitro models
Language English
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PublicationTitle Biofabrication
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Snippet Three-dimensional (3D) cell culture has tremendous advantages to closely mimic the architecture and microenvironment of healthy tissue and organs, as well as...
Abstract Three-dimensional (3D) cell culture has tremendous advantages to closely mimic the in vivo architecture and microenvironment of healthy tissue and...
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SubjectTerms Bioprinting
Cell Culture Techniques
Hydrogels
Spheroids, Cellular
Tissue Engineering
Title Cell spheroids as a versatile research platform: formation mechanisms, high throughput production, characterization and applications
URI https://www.ncbi.nlm.nih.gov/pubmed/33592595
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