A novel model to culture cells from giant cell tumor of bone using three‐dimensional (3D) polycaprolactone scaffold

Two‐dimensional (2D) culture of cells from giant cell tumor of bone (GCTB) is affected by loss of the multinucleated giant cells in subsequent passages. Therefore, there is limited time to study GCTB with all its histological components in 2D culture. Here, we explored the possibility of culturing G...

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Published inEngineering in life sciences Vol. 21; no. 8-9; pp. 539 - 543
Main Authors Estrada‐Villaseñor, Eréndira, Valdés‐Flores, Margarita, Meneses‐García, Abelardo, Silva‐Bermudez, Phaedra, Pichardo‐Bahena, Raul, Ostoa‐Saloma, Pedro, Mercado‐Celis, Gabriela, Delgado‐Cedillo, Ernesto D., Olivos‐Meza, Anell, Landa‐Solís, Carlos
Format Journal Article
LanguageEnglish
Published Germany John Wiley & Sons, Inc 01.09.2021
John Wiley and Sons Inc
Wiley-VCH
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Online AccessGet full text
ISSN1618-0240
1618-2863
DOI10.1002/elsc.202100020

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Abstract Two‐dimensional (2D) culture of cells from giant cell tumor of bone (GCTB) is affected by loss of the multinucleated giant cells in subsequent passages. Therefore, there is limited time to study GCTB with all its histological components in 2D culture. Here, we explored the possibility of culturing GCTB cells on a polycaprolactone (PCL)‐printed scaffold. We also evaluated the viability of the cultured cells and their adherence to the PCL scaffold at day 14 days using immunofluorescence analysis with calcein, vinculin, and phalloidin. Using the histological technique with hematoxylin and eosin staining, we observed all the histological components of GCTB in this 3D model. Immunohistochemical assays with cathepsin K, p63, and receptor activator of nuclear factor (NF)‐κB ligand (RANKL) yielded positive results in this construct, which allowed us to confirm that the seeded cells maintained the expression of GCTB markers. Based on these findings, we concluded that the PCL scaffold is an efficient model to culture GCTB cells, and the cell viability and adherence to the scaffold can be preserved for up to 14 days. Moreover, this model can also be used in subsequent studies to assess in vitro cell–cell interactions and antineoplastic efficacy of certain agents to establish a treatment against GCTB.
AbstractList Abstract Two‐dimensional (2D) culture of cells from giant cell tumor of bone (GCTB) is affected by loss of the multinucleated giant cells in subsequent passages. Therefore, there is limited time to study GCTB with all its histological components in 2D culture. Here, we explored the possibility of culturing GCTB cells on a polycaprolactone (PCL)‐printed scaffold. We also evaluated the viability of the cultured cells and their adherence to the PCL scaffold at day 14 days using immunofluorescence analysis with calcein, vinculin, and phalloidin. Using the histological technique with hematoxylin and eosin staining, we observed all the histological components of GCTB in this 3D model. Immunohistochemical assays with cathepsin K, p63, and receptor activator of nuclear factor (NF)‐κB ligand (RANKL) yielded positive results in this construct, which allowed us to confirm that the seeded cells maintained the expression of GCTB markers. Based on these findings, we concluded that the PCL scaffold is an efficient model to culture GCTB cells, and the cell viability and adherence to the scaffold can be preserved for up to 14 days. Moreover, this model can also be used in subsequent studies to assess in vitro cell–cell interactions and antineoplastic efficacy of certain agents to establish a treatment against GCTB.
Two-dimensional (2D) culture of cells from giant cell tumor of bone (GCTB) is affected by loss of the multinucleated giant cells in subsequent passages. Therefore, there is limited time to study GCTB with all its histological components in 2D culture. Here, we explored the possibility of culturing GCTB cells on a polycaprolactone (PCL)-printed scaffold. We also evaluated the viability of the cultured cells and their adherence to the PCL scaffold at day 14 days using immunofluorescence analysis with calcein, vinculin, and phalloidin. Using the histological technique with hematoxylin and eosin staining, we observed all the histological components of GCTB in this 3D model. Immunohistochemical assays with cathepsin K, p63, and receptor activator of nuclear factor (NF)-κB ligand (RANKL) yielded positive results in this construct, which allowed us to confirm that the seeded cells maintained the expression of GCTB markers. Based on these findings, we concluded that the PCL scaffold is an efficient model to culture GCTB cells, and the cell viability and adherence to the scaffold can be preserved for up to 14 days. Moreover, this model can also be used in subsequent studies to assess in vitro cell-cell interactions and antineoplastic efficacy of certain agents to establish a treatment against GCTB.Two-dimensional (2D) culture of cells from giant cell tumor of bone (GCTB) is affected by loss of the multinucleated giant cells in subsequent passages. Therefore, there is limited time to study GCTB with all its histological components in 2D culture. Here, we explored the possibility of culturing GCTB cells on a polycaprolactone (PCL)-printed scaffold. We also evaluated the viability of the cultured cells and their adherence to the PCL scaffold at day 14 days using immunofluorescence analysis with calcein, vinculin, and phalloidin. Using the histological technique with hematoxylin and eosin staining, we observed all the histological components of GCTB in this 3D model. Immunohistochemical assays with cathepsin K, p63, and receptor activator of nuclear factor (NF)-κB ligand (RANKL) yielded positive results in this construct, which allowed us to confirm that the seeded cells maintained the expression of GCTB markers. Based on these findings, we concluded that the PCL scaffold is an efficient model to culture GCTB cells, and the cell viability and adherence to the scaffold can be preserved for up to 14 days. Moreover, this model can also be used in subsequent studies to assess in vitro cell-cell interactions and antineoplastic efficacy of certain agents to establish a treatment against GCTB.
Two-dimensional (2D) culture of cells from giant cell tumor of bone (GCTB) is affected by loss of the multinucleated giant cells in subsequent passages. Therefore, there is limited time to study GCTB with all its histological components in 2D culture. Here, we explored the possibility of culturing GCTB cells on a polycaprolactone (PCL)-printed scaffold. We also evaluated the viability of the cultured cells and their adherence to the PCL scaffold at day 14 days using immunofluorescence analysis with calcein, vinculin, and phalloidin. Using the histological technique with hematoxylin and eosin staining, we observed all the histological components of GCTB in this 3D model. Immunohistochemical assays with cathepsin K, p63, and receptor activator of nuclear factor (NF)-κB ligand (RANKL) yielded positive results in this construct, which allowed us to confirm that the seeded cells maintained the expression of GCTB markers. Based on these findings, we concluded that the PCL scaffold is an efficient model to culture GCTB cells, and the cell viability and adherence to the scaffold can be preserved for up to 14 days. Moreover, this model can also be used in subsequent studies to assess in vitro cell-cell interactions and antineoplastic efficacy of certain agents to establish a treatment against GCTB.
Audience Academic
Author Meneses‐García, Abelardo
Landa‐Solís, Carlos
Estrada‐Villaseñor, Eréndira
Valdés‐Flores, Margarita
Pichardo‐Bahena, Raul
Delgado‐Cedillo, Ernesto D.
Silva‐Bermudez, Phaedra
Ostoa‐Saloma, Pedro
Olivos‐Meza, Anell
Mercado‐Celis, Gabriela
AuthorAffiliation 1 Pathology Service National Institute of Rehabilitation Luis Guillermo Ibarra Ibarra Mexico City Mexico
5 Department of Immunology Institute of Biomedical Research National Autonomous University of Mexico Mexico City Mexico
6 Laboratory of Clinical Genomics Division of Graduate Studies and Research Faculty of Odontology National Autonomous University of Mexico Mexico City Mexico
3 Pathology Service National Cancer Institute Mexico City Mexico
2 Genetics Laboratory National Institute of Rehabilitation Luis Guillermo Ibarra Ibarra Mexico City Mexico
4 Tissue Engineering and Cell Therapy Unit National Institute of Rehabilitation Luis Guillermo Ibarra Ibarra Mexico City Mexico
7 Bone Tumors Service National Institute of Rehabilitation Luis Guillermo Ibarra Ibarra Mexico City Mexico
8 Sports Orthopedics and Arthroscopy National Institute of Rehabilitation Luis Guillermo Ibarra Ibarra Mexico City Mexico
AuthorAffiliation_xml – name: 5 Department of Immunology Institute of Biomedical Research National Autonomous University of Mexico Mexico City Mexico
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Keywords 3D tumor models
3D PCL scaffold
giant cell tumor of bone
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  article-title: Characterization of cells cultured from human giant‐cell tumors of bone. Phenotypic relationship to the monocyte‐macrophage and osteoclast
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Snippet Two‐dimensional (2D) culture of cells from giant cell tumor of bone (GCTB) is affected by loss of the multinucleated giant cells in subsequent passages....
Two-dimensional (2D) culture of cells from giant cell tumor of bone (GCTB) is affected by loss of the multinucleated giant cells in subsequent passages....
Abstract Two‐dimensional (2D) culture of cells from giant cell tumor of bone (GCTB) is affected by loss of the multinucleated giant cells in subsequent...
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SubjectTerms 3D PCL scaffold
3D tumor models
Analysis
Calcein
Carbon dioxide
Cathepsin K
Cathepsins
Cell adhesion & migration
Cell culture
Cell interactions
Cell viability
Cytokines
Ewings sarcoma
giant cell tumor of bone
Giant cells
Immunofluorescence
Phalloidin
Polycaprolactone
Protein expression
Proteins
Scaffolds
Short Communication
Three dimensional models
TRANCE protein
Tumors
Vinculin
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Title A novel model to culture cells from giant cell tumor of bone using three‐dimensional (3D) polycaprolactone scaffold
URI https://onlinelibrary.wiley.com/doi/abs/10.1002%2Felsc.202100020
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