Purification and functional characterization of novel human skeletal stem cell lineages

Malachia Y. Hoover, Thomas H. Ambrosi, Holly M. Steininger, Lauren S. Koepke, Yuting Wang, Liming Zhao, Matthew P. Murphy, Alina A. Alam, Elizabeth J. Arouge, M. Gohazrua K. Butler, Eri Takematsu, Suzan P. Stavitsky, Serena Hu, Debashis Sahoo, Rahul Sinha, Maurizio Morri, Norma Neff, Julius Bishop, Michael Gardner, Stuart Goodman, Michael Longaker, Charles K. F. Chan

Published: 2023-06-13 DOI: 10.1038/s41596-023-00836-5

human skeletal stem cells

flow cytometric isolation

functional characterization

skeletally restricted lineages

single-cell RNA sequencing

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Human skeletal stem cells (hSSCs) hold tremendous therapeutic potential for developing new clinical strategies to effectively combat congenital and age-related musculoskeletal disorders. Unfortunately, refined methodologies for the proper isolation of bon
Introduction
Materials
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Timing
Anticipated results
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References
ACKNOWLEDGMENTS
ACKNOWLEDGMENTS

Ono, N., Balani, D. H. & Kronenberg, H. M. Stem and progenitor cells in skeletal development. Curr. Top. Dev. Biol. 133, 1–24 (2019).
Matsushita, Y., Ono, W. & Ono, N. Skeletal stem cells for bone development and repair: diversity matters. Curr. Osteoporos. Rep. 18, 189–198 (2020).
Ambrosi, T. H., Longaker, M. T. & Chan, C. K. F. A revised perspective of skeletal stem cell biology. Front. Cell. Dev. Biol. 13, 189 (2019).
Dzierzak, E. & Bigas, A. Blood development: hematopoietic stem cell dependence and independence. Cell Stem Cell 22, 639–651 (2018).
Haas, S., Trumpp, A. & Milsom, M. D. Causes and consequences of hematopoietic stem cell heterogeneity. Cell Stem Cell 22, 627–638 (2018).
Sakamaki, T. et al. Hoxb5 defines the heterogeneity of self-renewal capacity in the hematopoietic stem cell compartment. Biochem. Biophys. Res. Commun. 539, 34–41 (2021).
Pittenger, M. F. et al. Multilineage potential of adult human mesenchymal stem cells. Science 284, 143–147 (1999).
Jiang, Y. et al. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 418, 41–49 (2002).
Kagami, H., Agata, H. & Tojo, A. Bone marrow stromal cells (bone marrow-derived multipotent mesenchymal stromal cells) for bone tissue engineering: basic science to clinical translation. Int. J. Biochem. Cell Biol. 43, 286–289 (2011).
Friedenstein, A. J., Chailakhyan, R. K. & Gerasimov, U. V. Bone marrow osteogenic stem cells: in vitro cultivation and transplantation in diffusion chambers. Cell Tissue Kinet. 20, 263–272 (1987).
Friedenstein, A. J., Gorskaja, J. F. & Kulagina, N. N. Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp. Hematol. 4, 267–274 (1976).
Zhou, B. O., Yue, R., Murphy, M. M., Peyer, J. G. & Morrison, S. J. Leptin-receptor-expressing mesenchymal stromal cells represent the main source of bone formed by adult bone marrow. Cell Stem Cell 15, 154–168 (2014).
Hall, S. R. et al. Identification and isolation of small CD44-negative mesenchymal stem/progenitor cells from human bone marrow using elutriation and polychromatic flow cytometry. Stem Cells Transl. Med. 2, 567–578 (2013).
Huang, Y. et al. Characterization and immunogenicity of bone marrow-derived mesenchymal stem cells under osteoporotic conditions. Sci. China Life Sci. 63, 429–442 (2020).
Chan, C. K. et al. Identification and specification of the mouse skeletal stem cell. Cell 160, 285–298 (2015).
Chan, C. K. F. et al. Identification of the human skeletal stem cell. Cell 175, 43–56.e21 (2018).
Dominici, M. et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The international society for cellular therapy position statement. Cytotherapy 8, 315–317 (2006).
Chung, M. T. et al. CD90 (Thy-1)-positive selection enhances osteogenic capacity of human adipose-derived stromal cells. Tissue Eng. Part A 19, 989–997 (2013).
Lange, A., Dlubek, D., Drabczak-Skrzypek, D., Bogunia-Kubik, K. & Emilia Marrow cells cultured in MSC medium expand to CD73, CD90 and CD105 cells of fibroblast-like morphology. Blood 106, 4319–4319 (2005).
Quirici, N. et al. Isolation of bone marrow mesenchymal stem cells by anti-nerve growth factor receptor antibodies. Exp. Hematol. 30, 783–791 (2002).
Sorrentino, A. et al. Isolation and characterization of CD146+ multipotent mesenchymal stromal cells. Exp. Hematol. 36, 1035–1046 (2008).
Sacchetti, B. et al. No identical “mesenchymal stem cells” at different times and sites: human committed progenitors of distinct origin and differentiation potential are incorporated as adventitial cells in microvessels. Stem Cell Rep. 6, 897–913 (2016).
Arthur, A. & Gronthos, S. Clinical application of bone marrow mesenchymal stem/stromal cells to repair skeletal tissue. Int. J. Mol. Sci. 21, 9759 (2020).
Mohamed-Ahmed, S. et al. Comparison of bone regenerative capacity of donor-matched human adipose-derived and bone marrow mesenchymal stem cells. Cell Tissue Res. 383, 1061–1075 (2021).
Elefteriou, F. Impact of the autonomic nervous system on the skeleton. Physiol. Rev. 98, 1083–1112 (2018).
Raaijmakers, M. H. et al. Bone progenitor dysfunction induces myelodysplasia and secondary leukaemia. Nature 464, 852–7 (2010).
Fornetti, J., Welm, A. L. & Stewart, S. A. Understanding the bone in cancer metastasis. J. Bone Miner. Res. 33, 2099–2113 (2018).
Murphy, M. P. et al. Articular cartilage regeneration by activated skeletal stem cells. Nat. Med. 26, 1583–1592 (2020).
Esposito, M., Guise, T. & Kang, Y. The biology of bone metastasis. Cold Spring Harb. Perspect. Med. 8, a031252 (2018).
Marecic, O. et al. Identification and characterization of an injury-induced skeletal progenitor. Proc. Natl Acad. Sci. USA 112, 9920–9925 (2015).
Ambrosi, T. H. et al. Aged skeletal stem cells generate an inflammatory degenerative niche. Nature 597, 256–262 (2021).
Ambrosi, T. H. et al. Geriatric fragility fractures are associated with a human skeletal stem cell defect. Aging Cell 19, e13164 (2020).
Gulati, G. S. et al. Isolation and functional assessment of mouse skeletal stem cell lineage. Nat. Protoc. 13, 1294–1309 (2018).
Goodnough, L. H. Cross-species comparisons reveal resistance of human skeletal stem cells to inhibition by non-steroidal anti-inflammatory drugs. Front. Endocrinol. 13, 924927 (2022).
Ambrosi, T. H. et al. Distinct skeletal stem cell types orchestrate long bone skeletogenesis. eLife 10, e66063 (2021).
Morton, J. J. et al. Dual use of hematopoietic and mesenchymal stem cells enhances engraftment and immune cell trafficking in an allogeneic humanized mouse model of head and neck cancer. Mol. Carcinog. 57, 1651–1663 (2018).
Mian, S. A., Anjos-Afonso, F. & Bonnet, D. Advances in human immune system mouse models for studying human hematopoiesis and cancer immunotherapy. Front. Immunol. 2, 619236 (2021).
Jain, R. & Jain, P. C. Production and partial characterization of collagenase of Streptomyces exfoliatus CFS 1068 using poultry feather. Indian J. Exp. Biol. 48, 174–178 (2010).
Steinhardt, R. Rips repaired. Nature 436, 925 (2005).
Picelli, S. et al. Full-length RNA-seq from single cells using Smart-seq2. Nat. Protoc. 9, 171–181 (2014).
Wang, X., He, Y., Zhang, Q., Ren, X. & Zhang, Z. Direct comparative analyses of 10X Genomics chromium and Smart-seq2. Genomics Proteom. Bioinforma. 19, 253–266 (2021).
Ziegenhain, C. et al. Comparative analysis of single-cell RNA sequencing methods. Mol. Cell 65, 631–643.e4 (2017).

Purification and functional characterization of novel human skeletal stem cell lineages

Extended

Extended Data Fig. 1 FACS gating strategy for other hSSC sources.

Extended Data Fig. 2 FACS gating strategy for SSCs of bone marrow reamings.

Extended Data Fig. 3 Fluorescence Minus One (FMO) gating strategy.

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