Journal: Nature Communications
Article Title: Functional and analytical recapitulation of osteoclast biology on demineralized bone paper
doi: 10.1038/s41467-023-44000-9
Figure Lengend Snippet: A Schematic of human bone marrow stromal cells (hBMSCs) isolation, expansion, and differentiation into human osteoblasts (hOBs) on DBP. After 2 weeks of remineralization in osteogenic differentiation medium, humanized RdBP was prepared by decellularization. B (i) hOBs adhere following the collagen fiber orientation on DBP but randomly on TCP. (ii) Alizarin red mineral staining after hOB culture on DBP and TCP. (iii) Time-course quantitative measurement of mineral deposition by fluorescent calcein staining ( n = 6 independent samples). C (i) Schematic of human CD14 + mononuclear cells (hCD14 + MNCs) isolation and expansion from peripheral blood using magnetic-activated cell sorting system. (ii) Brightfield images of ex vivo expanded hCD14 + cells for 1 week culture with hM-CSF. Representative images from 3 independent experiments. (iii) hCD14 + cells cultured on TCP, HP, and RdBP for 2 weeks with hM-CSF and hRANKL to induce hOC differentiation. D (i) Representative images of hOCs on TCP, HP, and RdBP. (green: phalloidin, blue: DAPI) (ii) Characterization of hOC sizes on TCP, HP, and RdBP after 2 weeks of the stimulated culture ( n = 20 cells from three independent experiments). E (i) Representative time-course fluorescent images of calcein-coated mineral resorption by hOCs on control, OS680-, and Zoledronate (Zol)-treated RdBP. (ii) Time-course quantitative monitoring of resorption areas. ( n = 3 independent samples) Inner panel images show mineral resorption (purple: OS680). F (i) Representative fluorescent images of hOCs on control, OS680-, and Zol-treated RdBP (green: phalloidin, blue: DAPI). (ii) Normalized hOC density at the end of 2 weeks culture ( n = 5 independent samples). The data are presented as mean ± standard deviation. P -values are derived from unpaired two-tailed t -tests. ns: not significant ( p > 0.05). Related source data are provided as a Source Data file.
Article Snippet: Fresh human bone marrow aspirate (50 mL) was purchased from Lonza (age 25–45 male and female donors).
Techniques: Isolation, Staining, FACS, Ex Vivo, Cell Culture, Control, Standard Deviation, Derivative Assay, Two Tailed Test
