Human skin-derived keratinocytes and fibroblasts co-cultured on 3D poly ε-caprolactone scaffold support in vitro HSC differentiation into T-lineage committed cells

Loredana Palamaro, Vincenzo Guarino, Giulia Scalia, Dario Antonini, Luigia De Falco, Gabriella Bianchino, Anna Fusco, Rosa Romano, Vitina Grieco, Caterina Missero, Luigi Del Vecchio, Luigi Ambrosio, Claudio Pignata

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)703-714
Number of pages12
JournalInternational Immunology
Volume25
Issue number12
DOIs
Publication statusPublished - Dec 2013

Keywords

  • HSC differentiation
  • PCL scaffold
  • Skin
  • T-cell development
  • Thymopoiesis

ASJC Scopus subject areas

  • Immunology

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