Abstract
The microenvironment plays a pivotal role for cell survival and functional regulation, and directs the cell fate determination. The biological functions of DCs have been extensively investigated to date. However, the influences of the microenvironment on the differentiation of bone marrow cells (BMCs) into dendritic cells (DCs) are not well defined. Here, we established a 3D collagen scaffold microenvironment to investigate whether such 3D collagen scaffolds could provide a favourable niche for BMCs to differentiate into specialised DCs. We found that BMCs embedded in the 3D collagen scaffold differentiated into a distinct subset of DC, exhibiting high expression of CD11b and low expression of CD11c, co-stimulator (CD40, CD80, CD83, and CD86) and MHC-II molecules compared to those grown in 2D culture. DCs cultured in the 3D collagen scaffold possessed weak antigen uptake ability and inhibited T-cell proliferation in vitro; in addition, they exhibited potent immunoregulatory function to alleviate allo-delay type hypersensitivity when transferred in vivo. Thus, DCs differentiated in the 3D collagen scaffold were defined as regulatory DCs, indicating that collagen scaffold microenvironments probably play an important role in modulating the lineage commitment of DCs and therefore might be applied as a promising tool for generation of specialised DCs.
Highlights
Demonstrated that 3D cultured cells exhibit considerable differences in cellular morphology, phenotype, and biological functions compared to cells cultured under 2D conditions[14,15,16,17]
We cultured bone marrow cells (BMCs) in 3D collagen scaffolds to facilitate their differentiation into a specific subset of Dendritic cells (DCs), DCregs, and evaluated their functional characteristics
DCs cultured in 3D collagen scaffolds exhibited low expression of MHC and co-stimulatory molecules (CD40, CD80, CD83, and CD86) and demonstrated immunoregulatory functions both in vitro and in vivo
Summary
Demonstrated that 3D cultured cells exhibit considerable differences in cellular morphology, phenotype, and biological functions compared to cells cultured under 2D conditions[14,15,16,17]. Our previous studies have shown that 3D culture was able to maintain the pluripotency of stem cells and direct the cell fate determination of mouse embryonic stem cells[14,15]. We sought to explore the role of collagen scaffold microenvironments in the differentiation of bone marrow cells (BMCs) into DCs. We established an in vitro 3D collagen scaffold microenvironment and investigated whether BMCs in this culture system demonstrated the ability to differentiate into highly specialised populations of DCs
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