title: Human skin-derived keratinocytes and fibroblasts co-cultured on 3D poly ε-caprolactone scaffold support in vitro HSC differentiation into T-lineage committed cells.
abstract: In humans, the thymus is the primary lymphoid organ able to support the development of T cells through its three-dimensional (3D) organization of the thymic stromal cells. Since a remarkable number of similarities are shared between the thymic epithelial cells (TECs) and skin-derived keratinocytes and fibroblasts, in this study we used human keratinocytes seeded with fibroblasts on the 3D poly ε-caprolactone scaffold to evaluate their ability to replace TECs in supporting T-cell differentiation from human haematopoietic stem cells (HSCs). We observed that in the multicellular biocomposite, early thymocytes expressing CD7(+)CD1a(+), peculiar markers of an initial T-cell commitment, were de novo generated. Molecular studies of genes selectively expressed during T-cell development revealed that TAL1 was down-regulated and Spi-B was up-regulated in the cell suspension, consistently with a T-cell lineage commitment. Moreover, PTCRA and RAG2 expression was detected, indicative of a recombinant activity, required for the generation of a T-cell receptor repertoire. Our results indicate that in the multicellular biocomposite, containing skin-derived elements in the absence of thymic stroma, HSCs do start differentiating toward a T-cell lineage commitment. In conclusion, the construct described in this study exerts some properties of a lymphoid organoid, suitable for future clinical applications in cell-based therapies.
authors.affiliation: Department of Translational Medical Sciences, 'Federico II' University, Naples 80131, Italy.
authors.forename: Loredana
authors.initials: L
authors.lastname: Palamaro
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Guarino
authors.forename: Giulia
authors.initials: G
authors.lastname: Scalia
authors.forename: Dario
authors.initials: D
authors.lastname: Antonini
authors.forename: Luigia
authors.initials: L
authors.lastname: De Falco
authors.forename: Gabriella
authors.initials: G
authors.lastname: Bianchino
authors.forename: Anna
authors.initials: A
authors.lastname: Fusco
authors.forename: Rosa
authors.initials: R
authors.lastname: Romano
authors.forename: Vitina
authors.initials: V
authors.lastname: Grieco
authors.forename: Caterina
authors.initials: C
authors.lastname: Missero
authors.forename: Luigi
authors.initials: L
authors.lastname: Del Vecchio
authors.forename: Luigi
authors.initials: L
authors.lastname: Ambrosio
authors.forename: Claudio
authors.initials: C
authors.lastname: Pignata
mesh_terms: D002208:Caproates / Q000737:chemistry; D002450:Cell Communication; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D018920:Coculture Techniques; D000078404:Epidermal Cells; D005347:Fibroblasts / Q000502:physiology* / Q000648:ultrastructure; D005786:Gene Expression Regulation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D015603:Keratinocytes / Q000502:physiology* / Q000648:ultrastructure; D007783:Lactones / Q000737:chemistry; D007963:Leukocytes, Mononuclear / Q000502:physiology; D010641:Phenotype; D016062:Porosity; D054504:Precursor Cells, T-Lymphoid / Q000166:cytology* / Q000378:metabolism; D013950:Thymus Gland / Q000166:cytology / Q000502:physiology; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D002208:Caproates; D007783:Lactones; C121056:caprolactone
keywords: HSC differentiation; PCL scaffold; T-cell development; skin; thymopoiesis