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Generation of Organotypic Multicellular Spheres by Magnetic Levitation: Model for the Study of Human Hematopoietic Stem Cells Microenvironment

BACKGROUND AND OBJECTIVE: The characteristics of human hematopoietic stem cells are conditioned by the microenvironment of the bone marrow, where they interact with other cell populations, such as mesenchymal stem cells and endothelial cells; however, the study of this microenvironment is complex. T...

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Autores principales: Mejía-Cruz, Claudia Camila, Barreto-Durán, Emilia, Pardo-Pérez, María Alejandra, Jimenez, María Camila, Rincón, Julieth, Vanegas, Karen, Rodríguez, Jorge Luis, Jaramillo-Garcia, Luis Fernando, Ulloa, Juan Carlos, Díaz, Rodolfo Martínez, Leal-García, Efrain, Pérez-Núñez, Rafael, Barreto, Alfonso, Rodríguez-Pardo, Viviana M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Korean Society for Stem Cell Research 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6457696/
https://www.ncbi.nlm.nih.gov/pubmed/30836729
http://dx.doi.org/10.15283/ijsc18061
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author Mejía-Cruz, Claudia Camila
Barreto-Durán, Emilia
Pardo-Pérez, María Alejandra
Jimenez, María Camila
Rincón, Julieth
Vanegas, Karen
Rodríguez, Jorge Luis
Jaramillo-Garcia, Luis Fernando
Ulloa, Juan Carlos
Díaz, Rodolfo Martínez
Leal-García, Efrain
Pérez-Núñez, Rafael
Barreto, Alfonso
Rodríguez-Pardo, Viviana M
author_facet Mejía-Cruz, Claudia Camila
Barreto-Durán, Emilia
Pardo-Pérez, María Alejandra
Jimenez, María Camila
Rincón, Julieth
Vanegas, Karen
Rodríguez, Jorge Luis
Jaramillo-Garcia, Luis Fernando
Ulloa, Juan Carlos
Díaz, Rodolfo Martínez
Leal-García, Efrain
Pérez-Núñez, Rafael
Barreto, Alfonso
Rodríguez-Pardo, Viviana M
author_sort Mejía-Cruz, Claudia Camila
collection PubMed
description BACKGROUND AND OBJECTIVE: The characteristics of human hematopoietic stem cells are conditioned by the microenvironment of the bone marrow, where they interact with other cell populations, such as mesenchymal stem cells and endothelial cells; however, the study of this microenvironment is complex. The objective of this work was to develop a 3D culture system by magnetic levitation that imitates the microenvironment of human HSC. METHODS AND RESULTS: Human bone marrow-mesenchymal stem cells, umbilical cord blood-hematopoietic stem cells and a non-tumoral endothelial cell line (CC2811, Lonza(®)) were used to develop organotypic multicellular spheres by the magnetic levitation method. We obtained viable structures with an average sphericity index greater than 0.6, an average volume of 0.5 mm(3) and a percentage of aggregation greater than 70%. Histological studies of the organotypic multicellular spheres used hematoxylin and eosin stains, and an evaluation of vimentin expression by means of immunohistochemistry demonstrated an organized internal structure without picnotic cells and a high expression of vimentin. The functional capacity of human hematopoietic stem cells after organotypic multicellular spheres culture was evaluated by multipotency tests, and it was demonstrated that 3D structures without exogenous Flt3L are autonomous in the maintenance of multipotency of human hematopoietic stem cells. CONCLUSIONS: We developed organotypic multicellular spheres from normal human cells that mimic the microenvironment of the human hematopoietic stem cells. These structures are the prototype for the development of complex organoids that allow the further study of the biology of normal human stem cells and their potential in regenerative medicine.
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spelling pubmed-64576962019-04-19 Generation of Organotypic Multicellular Spheres by Magnetic Levitation: Model for the Study of Human Hematopoietic Stem Cells Microenvironment Mejía-Cruz, Claudia Camila Barreto-Durán, Emilia Pardo-Pérez, María Alejandra Jimenez, María Camila Rincón, Julieth Vanegas, Karen Rodríguez, Jorge Luis Jaramillo-Garcia, Luis Fernando Ulloa, Juan Carlos Díaz, Rodolfo Martínez Leal-García, Efrain Pérez-Núñez, Rafael Barreto, Alfonso Rodríguez-Pardo, Viviana M Int J Stem Cells Original Article BACKGROUND AND OBJECTIVE: The characteristics of human hematopoietic stem cells are conditioned by the microenvironment of the bone marrow, where they interact with other cell populations, such as mesenchymal stem cells and endothelial cells; however, the study of this microenvironment is complex. The objective of this work was to develop a 3D culture system by magnetic levitation that imitates the microenvironment of human HSC. METHODS AND RESULTS: Human bone marrow-mesenchymal stem cells, umbilical cord blood-hematopoietic stem cells and a non-tumoral endothelial cell line (CC2811, Lonza(®)) were used to develop organotypic multicellular spheres by the magnetic levitation method. We obtained viable structures with an average sphericity index greater than 0.6, an average volume of 0.5 mm(3) and a percentage of aggregation greater than 70%. Histological studies of the organotypic multicellular spheres used hematoxylin and eosin stains, and an evaluation of vimentin expression by means of immunohistochemistry demonstrated an organized internal structure without picnotic cells and a high expression of vimentin. The functional capacity of human hematopoietic stem cells after organotypic multicellular spheres culture was evaluated by multipotency tests, and it was demonstrated that 3D structures without exogenous Flt3L are autonomous in the maintenance of multipotency of human hematopoietic stem cells. CONCLUSIONS: We developed organotypic multicellular spheres from normal human cells that mimic the microenvironment of the human hematopoietic stem cells. These structures are the prototype for the development of complex organoids that allow the further study of the biology of normal human stem cells and their potential in regenerative medicine. Korean Society for Stem Cell Research 2019-02-28 /pmc/articles/PMC6457696/ /pubmed/30836729 http://dx.doi.org/10.15283/ijsc18061 Text en Copyright © 2019 by the Korean Society for Stem Cell Research This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Mejía-Cruz, Claudia Camila
Barreto-Durán, Emilia
Pardo-Pérez, María Alejandra
Jimenez, María Camila
Rincón, Julieth
Vanegas, Karen
Rodríguez, Jorge Luis
Jaramillo-Garcia, Luis Fernando
Ulloa, Juan Carlos
Díaz, Rodolfo Martínez
Leal-García, Efrain
Pérez-Núñez, Rafael
Barreto, Alfonso
Rodríguez-Pardo, Viviana M
Generation of Organotypic Multicellular Spheres by Magnetic Levitation: Model for the Study of Human Hematopoietic Stem Cells Microenvironment
title Generation of Organotypic Multicellular Spheres by Magnetic Levitation: Model for the Study of Human Hematopoietic Stem Cells Microenvironment
title_full Generation of Organotypic Multicellular Spheres by Magnetic Levitation: Model for the Study of Human Hematopoietic Stem Cells Microenvironment
title_fullStr Generation of Organotypic Multicellular Spheres by Magnetic Levitation: Model for the Study of Human Hematopoietic Stem Cells Microenvironment
title_full_unstemmed Generation of Organotypic Multicellular Spheres by Magnetic Levitation: Model for the Study of Human Hematopoietic Stem Cells Microenvironment
title_short Generation of Organotypic Multicellular Spheres by Magnetic Levitation: Model for the Study of Human Hematopoietic Stem Cells Microenvironment
title_sort generation of organotypic multicellular spheres by magnetic levitation: model for the study of human hematopoietic stem cells microenvironment
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6457696/
https://www.ncbi.nlm.nih.gov/pubmed/30836729
http://dx.doi.org/10.15283/ijsc18061
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