Osteoblasts and bone marrow mesenchymal stromal cells control hematopoietic stem cell migration and proliferation in 3D in vitro model

• Published in: PloS one Vol. 5; no. 2; p. e9093
• Main Authors: de Barros, Ana Paula D N, Takiya, Christina M, Garzoni, Luciana R, Leal-Ferreira, Mona Lisa, Dutra, Hélio S, Chiarini, Luciana B, Meirelles, Maria Nazareth, Borojevic, Radovan, Rossi, Maria Isabel D
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 08.02.2010; Public Library of Science (PLoS)
• Subjects: Active control; Adult; Analysis; Animals; Antigens, CD34 - analysis; Arthritis; Biocompatibility; Biomedical materials; Bone marrow; Bone Marrow Cells - cytology; Bone Marrow Cells - metabolism; CD34 antigen; Cell Biology; Cell Biology/Cell Adhesion; Cell Biology/Cell Growth and Division; Cell Culture Techniques; Cell Line; Cell migration; Cell Movement; Cell Proliferation; Cells (biology); Cells, Cultured; Chemokine CXCL12 - genetics; Chemokines; Coculture Techniques; Collagen; Colony-forming cells; Cord blood; CXCL12 protein; Diabetes; Extracellular matrix; Fetal Blood - cytology; Fibroblasts; Fibronectin; Fibronectins; Flow Cytometry; Gene expression; Gene Expression Profiling; Growth factors; Hematology/Hematopoiesis; Hematopoietic stem cells; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - metabolism; Humans; Hypoxia; Infant, Newborn; Laminin; Mesenchymal stem cells; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; Mesenchyme; Microscopy; Microscopy, Electron; mRNA; Osteoblasts; Osteoblasts - cytology; Osteoblasts - metabolism; Osteopontin; Progenitor cells; Proteins; Reverse Transcriptase Polymerase Chain Reaction; Rodents; Self-assembly; Spheroids; Spheroids, Cellular - cytology; Spheroids, Cellular - metabolism; Spheroids, Cellular - ultrastructure; Stem cells; Stromal cells; Stromal Cells - cytology; Stromal Cells - metabolism; Three dimensional models; Vascular endothelial growth factor; Wnt protein; Brazil; Rio de Janeiro Brazil
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0009093

Migration, proliferation, and differentiation of hematopoietic stem cells (HSCs) are dependent upon a complex three-dimensional (3D) bone marrow microenvironment. Although osteoblasts control the HSC pool, the subendosteal niche is complex and its cellular composition and the role of each cell population in HSC fate have not been established. In vivo models are complex and involve subtle species-specific differences, while bidimensional cultures do not reflect the 3D tissue organization. The aim of this study was to investigate in vitro the role of human bone marrow-derived mesenchymal stromal cells (BMSC) and active osteoblasts in control of migration, lodgment, and proliferation of HSCs. A complex mixed multicellular spheroid in vitro model was developed with human BMSC, undifferentiated or induced for one week into osteoblasts. A clear limit between the two stromal cells was established, and deposition of extracellular matrix proteins fibronectin, collagens I and IV, laminin, and osteopontin was similar to the observed in vivo. Noninduced BMSC cultured as spheroid expressed higher levels of mRNA for the chemokine CXCL12, and the growth factors Wnt5a and Kit ligand. Cord blood and bone marrow CD34(+) cells moved in and out the spheroids, and some lodged at the interface of the two stromal cells. Myeloid colony-forming cells were maintained after seven days of coculture with mixed spheroids, and the frequency of cycling CD34(+) cells was decreased. Undifferentiated and one-week osteo-induced BMSC self-assembled in a 3D spheroid and formed a microenvironment that is informative for hematopoietic progenitor cells, allowing their lodgment and controlling their proliferation.