Effect of donor variation on osteogenesis and vasculogenesis in hydrogel cocultures

Abstract

To introduce a functional vascular network into tissue‐engineered bone equivalents, human endothelial colony forming cells (ECFCs) and multipotent mesenchymal stromal cells (MSCs) can be cocultured. Here, we studied the impact of donor variation of human bone marrow‐derived MSCs and cord blood‐derived ECFCs on vasculogenesis and osteogenesis using a 3D in vitro coculture model. Further, to make the step towards cocultures consisting of cells derived from a single donor, we tested how induced pluripotent stem cell (iPSC)‐derived human endothelial cells (iECs) performed in coculture models. Cocultures with varying combinations of human donors of MSCs, ECFCs, or iECs were prepared in Matrigel. The constructs were cultured in an osteogenic differentiation medium. Following a 10‐day culture period, the length of the prevascular structures and osteogenic differentiation were evaluated for up to 21 days of culture. The particular combination of MSC and ECFC donors influenced the vasculogenic properties significantly and induced variation in osteogenic potential. In addition, the use of iECs in the cocultures resulted in prevascular structure formation in osteogenically differentiated constructs. Together, these results showed that close attention to the source of primary cells, such as ECFCs and MSCs, is critical to address variability in vasculogenic and osteogenic potential. The 3D coculture model appeared to successfully generate prevascularized constructs and were sufficient in exceeding the ~200 μm diffusion limit. In addition, iPSC‐derived cell lineages may decrease variability by providing a larger and potentially more uniform source of cells for future preclinical and clinical applications.

Use of Therapeutic Pathogen Recognition Receptor Ligands for Osteo-Immunomodulation

“…hMSCs were isolated and cryopreserved as described in detail previously, as this method of MSC isolation yields multipotent cells as shown by standard differentiation assays along osteogenic, adipogenic, and chondrogenic lineages [ 35 ]. Cells below passage 7 were used for the experiments.…”

Antibacterial Titanium Implants Biofunctionalized by Plasma Electrolytic Oxidation with Silver, Zinc, and Copper: A Systematic Review

“…However, their variability is high. Cells from multiple donors, therefore, need to be tested [ 185 ]. Cell lines, on the other hand, are homogenous and stable, while exhibiting little variability.…”

Substantial Overview on Mesenchymal Stem Cell Biological and Physical Properties as an Opportunity in Translational Medicine

“…We should also consider that the stromal tissue origin determines several biological properties of MSC [77,128], including either a dissimilar balance and expression of pro- and suppressive-angiogenic factors (HGF, IL-10, TGF-β1, INF-γ, trombospondin-1) [129,130] and variation of endothelial cell differentiation and angiogenic potential according to donors [131]. For instance, Ad-MSC are more efficient to sustain vascularisation than the stromal counterpart obtained from umbilical and endometrial tissues [130].…”