Versatile humanized niche model enables study of normal and malignant human hematopoiesis
Ander Abarrategi, … , Ghulam Mufti, Dominique Bonnet
Ander Abarrategi, … , Ghulam Mufti, Dominique Bonnet
Published January 9, 2017
Citation Information: J Clin Invest. 2017;127(2):543-548. https://doi.org/10.1172/JCI89364.
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Versatile humanized niche model enables study of normal and malignant human hematopoiesis

Abstract

The BM niche comprises a tightly controlled microenvironment formed by specific tissue and cells that regulates the behavior of hematopoietic stem cells (HSCs). Here, we have provided a 3D model that is tunable in different BM niche components and useful, both in vitro and in vivo, for studying the maintenance of normal and malignant hematopoiesis. Using scaffolds, we tested the capacity of different stromal cell types to support human HSCs. Scaffolds coated with human mesenchymal stromal cells (hMSCs) proved to be superior in terms of HSC engraftment and long-term maintenance when implanted in vivo. Moreover, we found that hMSC-coated scaffolds can be modulated to form humanized bone tissue, which was also able to support human HSC engraftment. Importantly, hMSC-coated humanized scaffolds were able to support the growth of leukemia patient cells in vivo, including the growth of samples that would not engraft the BM of immunodeficient mice. These results demonstrate that an s.c. implantation approach in a 3D carrier scaffold seeded with stromal cells is an effective in vivo niche model for studying human hematopoiesis. The various niche components of this model can be changed depending on the context to improve the engraftment of nonengrafting acute myeloid leukemia (AML) samples.

Authors

Ander Abarrategi, Katie Foster, Ashley Hamilton, Syed A. Mian, Diana Passaro, John Gribben, Ghulam Mufti, Dominique Bonnet

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Figure 1

UCB-derived HSPC engraftment in s.c. scaffolds.

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UCB-derived HSPC engraftment in s.c. scaffolds. (A and B) Comparative as...
(A and B) Comparative assay using a single hMSC donor and multiple UCB donors. Each point represents 1 mouse seeded with 6 scaffolds. For each UCB donor, 3 to 8 mice were transplanted. (A) hCD45 scaffold-engraftment level. D1-D5: different UCB Donors. (B) Lineage distribution of engrafted cells. (C and D) A comparative assay using a single UCB donor and multiple hMSC donors. Each point represents 1 mouse seeded with 6 scaffolds. For each hMSC donor, 3 to 6 mice were transplanted. (C) hCD45 scaffold–engraftment levels. For comparative purposes, the engraftment levels in BM of i.v.-injected mice are provided. Tukey’s test for multiple comparisons was applied. *P < 0.05. (D) Lineage distribution of engrafted cells. (E) Gross morphology of harvested scaffold. (F) microCT study of a harvested scaffold. The scaffold is mainly soft tissue, with sporadic calcification spots. (G) Immunofluorescence image showing hCD45 cells in the scaffold and (H) osterix-positive (OSX+) and osterix-negative human stroma cells. (I) Schematic of preparation and implantation of bone-forming scaffolds. (J) Whole-mouse microCT showing the s.c. ossicle formation. (K) Gross morphology of the harvested ossicle. (L) microCT study of a harvested ossicle. (M and N) Immunofluorescence images showing (M) human vimentin+ (hVIM) mesenchymal cells and adipocytes and mature neovascularized (endomucin+, End) BM. (N) Trabecular bone formed inside ossicles. Dotted line shows delimited trabecular bone area. hVIM+/Osx+ cells are osteocytes or osteoblasts in the bone surface areas. (O) hCD45+ engraftment levels in the ossicle model tested with multiple UCB donors. Each point represents 1 mouse with 2 scaffolds. For each UCB donor, 2 to 4 mice were transplanted. Scale bars: 1 mm (E, F, K, and L); 20 μm (G, H and N); 50 μm (M). All data were harvested at 12 weeks after implantation.