| 研究概要 |
The main purpose of this research is to engineer a three-dimensional (3D) bone marrow niche microenvironment in vitro using micro-technologies. Initially, various micro-technology based 3D culture methods including spheroids, collagen cell beads, and cell fibers were explored. During this process, the cell fiber technology recently developed in this lab was determined to be the most suitable method to construct the 3D niche microenvironment for the proposed application. Using a microfluidic double co-axial device, adherent cells can be encapsulated into the core region of hydrogel core-shell fibers (diameter : 200-500 um ; length : meter scale). As proof-of-concept, multipotent DFAT (de-differentiated fat) cells that have the potential to differentiate into bone, cartilage, fat, and muscle cells were encapsulated into the core region of alginate shell fibers. Various extracellular matrix (ECM) proteins including collagen and fibrin at different concentrations were used to change the overall mechanical stiffness of the cellular microenvironment. An ECM mixture of collagen and fibrin was finally chosen as the optimized condition for long term DFAT cell fiber culture. Using the cell fiber technology, it is possible to engineering 3D cellular constructs into various shapes and control the cellular orientation at the micro-scale. Thus far, we achieved in engineering coiled spring shaped DFAT cell fiber constructs by inducing the cells to differentiate into the smooth muscle cell lineage. By changing the differentiation induction media, DFAT cell fibers can be differentiated into the osteoblast lineage. The established cell fiber 3D culture system would be very useful for the in vitro construction of the bone marrow niche microenvironment.
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| 今後の研究の推進方策 |
In the second year, the research will be focused on utilizing the established cell fiber system to construct 3D cellular niche in vitro. Instead of using DFAT cells, mesenchymal stem cells derived from bone marrow, or osteoblasts, endothelial cells, and cancer cells will be encapsulated into the core region of alginate shell fibers. The possibility of co-culturing these different cell fibers into a more complex 3D niche construct will be explored. Various 3D cellular constructs composed of different ratios of the co-culture cell types will be formed and the proliferation of the cancer cells will be observed.
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