Long-term in vitro expansion of hematopoietic stem cells (HSCs), while maintaining their functionality and multilineage differentiation potential, is still challenging. In this study, three-dimensional (3D) high-porosity hydroxyapatite (HA) foams have been designed to closely mimic the chemistry and physical structure of cancellous bone. Furthermore, calcium oxide was distributed in the HA ceramics to provide surface calcium ion release, hypothesizing that a local surface calcium gradient supports HSC localization and maintenance. Primary human HSCs and osteoblasts were cocultured for 6 weeks. Controls were cultured in two-dimensional dishes, while scaffold cultures were performed with calcium nitrate-infiltrated HA scaffolds and untreated HA scaffolds. Cells were analyzed for surface markers by flow cytometry, metabolic activity, and hematopoietic multilineage differentiation potential. The release of calcium into culture medium was also determined. The implementation of HA scaffolds had a positive effect on erythrocyte colony formation capacity of HSCs, with an increased osteoblast fraction observed when compared to control cultures without scaffolds. The presentation of scaffolds did not affect metabolic turnover when compared to control cultures. In conclusion, 3D open-porous HA scaffolds provide a bone-like structure and enable the long-term maintenance of primary HSCs.
Keywords: calcium ions; differentiation; hematopoietic stem cells; hydroxyapatite; long-term culture.