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Nanoengineered 3D culture substrate enables superior long-term and polyclonal engraftment of genetically engineered hematopoietic stem and progenitor cells

ABSTRACT: Ex vivo culture of hematopoietic stem and progenitor cells is crucial for gene therapy applications but inadvertently triggers detrimental cellular responses, potentially threatening clinical success. In this study, we employ a biocompatible 3D scaffold with cell-scale resolution to provide mechanical support during ex vivo manipulation. This innovative 3D culture system improves multi-lineage differentiation and engraftment capacity by leveraging mechanobiological control over nuclear morphology, cytoskeleton organization, metabolism, and DNA integrity. Notably, 3D culture enables efficient genetic engineering across multiple platforms, including long-range gene editing, base- and prime-editing, and lentiviral gene addition. Moreover, this scaffold increases the clonal output and the long-term persistence of genetically engineered cells in xenotransplantation experiments and outperforms traditional 2D culture in a clinical protocol for Wiskott-Aldrich Syndrome. Overall, we propose a transformative approach to enhance the efficacy and safety of hematopoietic stem cell-based gene therapies within emerging and pre-existing clinical workflows.

ORGANISM(S): Homo sapiens

PROVIDER: GSE280536 | GEO | 2025/12/09

REPOSITORIES: GEO

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Nanoengineered 3D culture substrate enables superior long-term and polyclonal engraftment of genetically engineered hematopoietic stem and progenitor cells

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