3D multicellular spheroid for the study of human hematopoietic stem cells : synergistic effect between oxygen levels, mesenchymal stromal cells and endothelial cells
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Datum
2021-06-30Autoren
Barreto Duran, EmiliaJaramillo García, Luis Fernando
Mejía Cruz, Claudia Camila
Leal García, Efraín
Barreto Prieto, Alfonso
Rodríguez Pardo, Viviana Marcela
Corporate Author(s)
Pontificia Universidad Javeriana. Facultad de Medicina. Departamento de Ortopedia y Traumatología
Pontificia Universidad Javeriana. Facultad de Medicina. Hospital Universitario San Ignacio
Typ
Artículo de revista
ISSN
1179-2736
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Introduction: The human bone marrow microenvironment is composed of biological,
chemical and physical factors that act in a synergistic way to modulate hematopoietic stem
cell biology, such as mesenchymal stromal cells (MSCs), endothelial cells (ECs) and low
oxygen levels; however, it is difficult to mimic this human microenvironment in vitro.
Methods: In this work, we developed 3D multicellular spheroid (3D-MS) for the study of
human hematopoietic stem cells (HSCs) with some components of perivascular niche. HSCs
were isolated from umbilical cord blood, MSCs were isolated from human bone marrow and
a microvasculature EC line (CC-2811, Lonza®) was used. For the formation of a 3D
structure, a magnetic levitation culture system was used. Cultures were maintained in 21%,
3% and 1% O2 for 15 days. Culture volume, sphericity index and cell viability were
determined. Also, human HSC proliferation, phenotype and production of reactive oxygen
species were evaluated.
Results: After 15 days, 3D-MS exhibited viability greater than 80%. Histology results
showed structures without necrotic centers, and higher cellular proliferation with 3% O2.
An increase in the expression of the CD34 antigen and other hematopoietic antigens were
observed to 1% O2 with MSCs plus ECs and low ROS levels.
Conclusion: These findings suggest that 3D-MS formed by MSCs, ECs and HSCs exposed
to low concentrations of oxygen (1–3% O2) modulate human HSC behavior and mimics
some features of the perivascular niche, which could reduce the use of animal models and
deepen the relationship between the microenvironment of HSC and human hematological
diseases development.
Verknüpfen Sie mit der Ressource
https://pubmed.ncbi.nlm.nih.gov/34234608/#affiliation-1Herkunft
Journal of Blood Medicine; Volumen 12 , Páginas 517 - 528 (2021)
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