TY - JOUR
T1 - Human skin-derived keratinocytes and fibroblasts co-cultured on 3D poly ε-caprolactone scaffold support in vitro HSC differentiation into T-lineage committed cells
AU - Palamaro, Loredana
AU - Guarino, Vincenzo
AU - Scalia, Giulia
AU - Antonini, Dario
AU - Falco, Luigia De
AU - Bianchino, Gabriella
AU - Fusco, Anna
AU - Romano, Rosa
AU - Grieco, Vitina
AU - Missero, Caterina
AU - Vecchio, Luigi Del
AU - Ambrosio, Luigi
AU - Pignata, Claudio
PY - 2013/12
Y1 - 2013/12
N2 - 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.
AB - 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.
KW - HSC differentiation
KW - PCL scaffold
KW - Skin
KW - T-cell development
KW - Thymopoiesis
UR - http://www.scopus.com/inward/record.url?scp=84883720032&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84883720032&partnerID=8YFLogxK
U2 - 10.1093/intimm/dxt035
DO - 10.1093/intimm/dxt035
M3 - Article
C2 - 24038600
AN - SCOPUS:84883720032
VL - 25
SP - 703
EP - 714
JO - International Immunology
JF - International Immunology
SN - 0953-8178
IS - 12
ER -