Figure 3.

Molecularly Engineered hyC Can Remodel into Humanized Bone Organs of Distinct Blood Compositions

(A) Experimental design for the generation of humanized ossicles (hOss). Engineered hyCs are implanted into immunocompromised animals. Mice are humanized 6 weeks later by intravenous transplantation of human CD34+ isolated from cord blood. Constructs are retrieved 6 weeks later for analysis.

(B and C) (B) VENUS and VENUS-SDF1α hyCs successfully remodeled into ossicles, as shown macroscopically by blood colonization and (C) by microtomographic scans revealing the formation of mature bone tissue. Ossicles were retrieved after 12 weeks in vivo. Scale bars, 0.5 mm.

(D) Humanized ossicles display a chimeric blood composition organized in “islets” of human hematopoieisis.

(E) SDF1α-overexpressing ossicles displayed higher frequencies of human blood populations, as assessed by flow cytometry. n ≥ 18 biological replicates from four independent experiments. ∗p < 0.5, using one-way ANOVA test. CMPs, common myeloid progenitors; HSCs, hematopoietic stem cells; HSPCs, hematopoietic stem and progenitor cells; MEPs, megakaryocyte-erythroid progenitors; MPPs, multipotent progenitors. Data are represented as mean ± SEM.