Directed differentiation of hematopoietic precursors and functional osteoclasts from human ES and iPS cells
| Autor: | In-Hyun Park, Aline Bozec, Agamemnon E. Grigoriadis, Erwin F. Wagner, Gudrun Stenbeck, Marion Kennedy, Gordon Keller, Fiona K Brunton |
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| Rok vydání: | 2010 |
| Předmět: |
Vascular Endothelial Growth Factor A
musculoskeletal diseases Macrophage colony-stimulating factor Pathology medicine.medical_specialty Hematopoiesis and Stem Cells Cellular differentiation Induced Pluripotent Stem Cells Immunology Osteoclasts Embryoid body Biology Biochemistry Cell Line 03 medical and health sciences 0302 clinical medicine Directed differentiation Osteoclast medicine Humans Induced pluripotent stem cell Embryonic Stem Cells 030304 developmental biology Myelopoiesis 0303 health sciences Macrophage Colony-Stimulating Factor RANK Ligand Cell Differentiation Cell Biology Hematology Hematopoietic Stem Cells Antigens Differentiation Embryonic stem cell Cell biology medicine.anatomical_structure Gene Expression Regulation 030220 oncology & carcinogenesis Stem cell |
| Zdroj: | Blood. 115:2769-2776 |
| ISSN: | 1528-0020 0006-4971 |
| DOI: | 10.1182/blood-2009-07-234690 |
| Popis: | The directed differentiation of human pluripotent stem cells offers the unique opportunity to generate a broad spectrum of human cell types and tissues for transplantation, drug discovery, and studying disease mechanisms. Here, we report the stepwise generation of bone-resorbing osteoclasts from human embryonic and induced pluripotent stem cells. Generation of a primitive streak-like population in embryoid bodies, followed by specification to hematopoiesis and myelopoiesis by vascular endothelial growth factor and hematopoietic cytokines in serum-free media, yielded a precursor population enriched for cells expressing the monocyte-macrophage lineage markers CD14, CD18, CD11b, and CD115. When plated in monolayer culture in the presence of macrophage colony-stimulating factor and receptor activator of nuclear factor-κB ligand (RANKL), these precursors formed large, multinucleated osteoclasts that expressed tartrate-resistant acid phosphatase and were capable of resorption. No tartrate-resistant acid phosphatase-positive multinucleated cells or resorption pits were observed in the absence of RANKL. Molecular analyses confirmed the expression of the osteoclast marker genes NFATc1, cathepsin K, and calcitonin receptor in a RANKL-dependent manner, and confocal microscopy demonstrated the coexpression of the αvβ3 integrin, cathepsin K and F-actin rings characteristic of active osteoclasts. Generating hematopoietic and osteoclast populations from human embryonic and induced pluripotent stem cells will be invaluable for understanding embryonic bone development and postnatal bone disease. |
| Databáze: | OpenAIRE |
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