Fabrication of bone marrow-like tissue
| Autor: | Kanae, Sayo, Shigehisa, Aoki, Nobuhiko, Kojima |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Aggregate
MC Methylcellulose BM Bone marrow 3D Three-dimensional PBS Phosphate buffered saline Methylcellulose DMEM Dulbecco's Modified Eagle Medium 2D Two-dimensional Three-dimensional culture HSCs Hematopoietic stem cells CXCL12 Chemokine (C-X-C motif) ligand 12 CAR cell CXCL12-abundant reticular cell PDGFRα Platelet-derived growth factor receptor alpha FBS Fetal bovine serum HE Hematoxylin-eosin Original Article Bone marrow Tissue engineering PFA Paraformaldehyde |
| Zdroj: | Regenerative Therapy |
| ISSN: | 2352-3204 |
| Popis: | A three-dimensional (3D) bone marrow (BM) culture system may facilitate research into the molecular mechanisms involved in hematopoiesis and BM diseases. However, because >90% of BM cells are composed of non-adherent blood cells, it is difficult to organize the dispersed BM cells into 3D multicellular spheroids using conventional aggregation methods such as hanging drop, and rotary shaking culture. The objective of this study was to reproduce BM-like tissue. We reported successful formation of BM aggregates using a 3% methylcellulose (MC) medium. This medium could aggregate even non-adherent materials. In MC medium, BM cells formed tissue-like aggregates within 24 h. Although the cell density of the BM-like tissue is slightly low, sections of the organoids resembled those of intact BM tissue. Cells of the BM-like tissue were approximately 70% viable after 7 days in culture. Staining for CD68, PDGFRα, and CXCL12 indicated that the BM-like tissue contained macrophages, and mesenchymal cells including CXCL12-abundant reticular cells. These results indicated that the method using MC medium effectively reconstitutes the BM-like tissue. Highlights • Dispersed-state BM cells can aggregate and organize in MC medium. • BM-like tissue was removed from MC medium after culturing 24 h. • BM-like tissue included macrophages, and PDGFRα+CXCL12+ cells. |
| Databáze: | OpenAIRE |
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