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T cells fail to develop in the human skin-cell explants system; an inconvenient truth

Meek, Bob, Van Elssen, Catharina H. M. J., Huijskens, Mirelle J. A. J., van der Stegen, Sjoukje J. C., Tonnaer, Siebe, Lumeij, Stijn B. J., Vanderlocht, Joris, Kirkland, Mark A., Hesselink, Reinout, Germeraad, Wilfred T. V. and Bos, Gerard M. J. 2011, T cells fail to develop in the human skin-cell explants system; an inconvenient truth, BMC immunology, vol. 12, pp. 1-13, doi: 10.1186/1471-2172-12-17.

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Title T cells fail to develop in the human skin-cell explants system; an inconvenient truth
Author(s) Meek, Bob
Van Elssen, Catharina H. M. J.
Huijskens, Mirelle J. A. J.
van der Stegen, Sjoukje J. C.
Tonnaer, Siebe
Lumeij, Stijn B. J.
Vanderlocht, Joris
Kirkland, Mark A.
Hesselink, Reinout
Germeraad, Wilfred T. V.
Bos, Gerard M. J.
Journal name BMC immunology
Volume number 12
Article ID 17
Start page 1
End page 13
Total pages 13
Publisher BioMed Central
Place of publication London, Eng.
Publication date 2011-02-18
ISSN 1471-2172
1471-2172
Keyword(s) Animals
Cell Culture Techniques
Cell Differentiation
Cell Line
Cells, Cultured
Coculture Techniques
Fibroblasts
Flow Cytometry
Fluorescent Antibody Technique
Hematopoietic Stem Cells
Humans
Intercellular Signaling Peptides and Proteins
Intracellular Signaling Peptides and Proteins
Keratinocytes
Membrane Proteins
Reverse Transcriptase Polymerase Chain Reaction
Skin
T-Lymphocytes
Summary BACKGROUND: Haplo-identical hematopoietic stem cell (HSC) transplantation is very successful in eradicating haematological tumours, but the long post-transplant T-lymphopenic phase is responsible for high morbidity and mortality rates. Clark et al. have described a skin-explant system capable of producing host-tolerant donor-HSC derived T-cells. Because this T-cell production platform has the potential to replenish the T-cell levels following transplantation, we set out to validate the skin-explant system.

RESULTS: Following the published procedures, while using the same commercial components, it was impossible to reproduce the skin-explant conditions required for HSC differentiation towards mature T-cells. The keratinocyte maturation procedure resulted in fragile cells with minimum expression of delta-like ligand (DLL). In most experiments the generated cells failed to adhere to carriers or were quickly outcompeted by fibroblasts. Consequently it was not possible to reproduce cell-culture conditions required for HSC differentiation into functional T-cells. Using cell-lines over-expressing DLL, we showed that the antibodies used by Clark et al. were unable to detect native DLL, but instead stained 7AAD+ cells. Therefore, it is unlikely that the observed T-lineage commitment from HSC is mediated by DLL expressed on keratinocytes. In addition, we did confirm expression of the Notch-ligand Jagged-1 by keratinocytes.

CONCLUSIONS: Currently, and unfortunately, it remains difficult to explain the development or growth of T-cells described by Clark et al., but for the fate of patients suffering from lymphopenia it is essential to both reproduce and understand how these co-cultures really "work". Fortunately, alternative procedures to speed-up T-cell reconstitution are being established and validated and may become available for patients in the near future.
Language eng
DOI 10.1186/1471-2172-12-17
Field of Research 110704 Cellular Immunology
1107 Immunology
Socio Economic Objective 970110 Expanding Knowledge in Technology
HERDC Research category C1.1 Refereed article in a scholarly journal
Grant ID Project: IS055002 to WTVG and GMJB
Copyright notice ©2011, The Authors
Free to Read? Yes
Use Rights Creative Commons Attribution licence
Persistent URL http://hdl.handle.net/10536/DRO/DU:30101100

Document type: Journal Article
Collections: Institute for Frontier Materials
Open Access Collection
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Every reasonable effort has been made to ensure that permission has been obtained for items included in DRO. If you believe that your rights have been infringed by this repository, please contact drosupport@deakin.edu.au.