Fibronectin and laminin enhance engraftibility of cultured hematopoietic stem cells

To test the hypothesis that extracellular matrix (ECM) components maintain stem cell property, murine bone marrow (BM) cells were expanded in fibronectin and laminin coated plate in the presence of cytokines. We observed significant phenotypic and functional improvement of expanded cells. In 10 days...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2006-12, Vol.350 (4), p.1000-1005
Main Authors: Sagar, Balla Murali Mohan, Rentala, Satyanarayana, Gopal, P.N.V., Sharma, Shalini, Mukhopadhyay, Asok
Format: Article
Language:English
Subjects:
Animals
Cell adhesion molecules
Cell Culture Techniques - methods
Cells, Cultured
Extracellular matrix
Fibronectins - chemistry
Fibronectins - pharmacology
Graft Survival - physiology
Hematopoietic Stem Cell Transplantation - methods
Hematopoietic stem cells
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - drug effects
Laminin - chemistry
Laminin - pharmacology
Mice
Mice, Inbred C57BL
Multi-lineage engraftibility
Primitiveness
Tissue Engineering - methods
Verapamil sensitivity
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Summary:To test the hypothesis that extracellular matrix (ECM) components maintain stem cell property, murine bone marrow (BM) cells were expanded in fibronectin and laminin coated plate in the presence of cytokines. We observed significant phenotypic and functional improvement of expanded cells. In 10 days, 800-fold expansion of colony-forming unit-granulocyte erythrocyte monocyte megakaryocyte (CFU-GEMM) was observed in the cultured cells. No apparent activation of cell cycle was observed, but CD29 and very late antigen-4 (VLA-4) expression was increased, as compared to the normal BM cells. A fraction of the expanded cells became verapamil sensitive, suggesting upregulation of multi-drug resistant gene(s), as found in the primitive hematopoietic stem cells (HSCs). Competitive repopulation assay confirmed that HSCs compartment was amplified during culture. Overall, our study clearly demonstrated that ex vivo culture of murine HSCs in the presence of fibronectin and laminin resulted in expansion of primitive stem cells and improvement in the marrow engraftibility.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2006.09.140