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Open Access 01-12-2011 | Correspondence

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

Authors: Bob Meek, Catharina HMJ Van Elssen, Mirelle JAJ Huijskens, Sjoukje JC van der Stegen, Siebe Tonnaer, Stijn BJ Lumeij, Joris Vanderlocht, Mark A Kirkland, Reinout Hesselink, Wilfred TV Germeraad, Gerard MJ Bos

Published in: BMC Immunology | Issue 1/2011

Abstract

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.

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Title: T cells fail to develop in the human skin-cell explants system; an inconvenient truth
Authors: Bob Meek
Catharina HMJ Van Elssen
Mirelle JAJ Huijskens
Sjoukje JC van der Stegen
Siebe Tonnaer
Stijn BJ Lumeij
Joris Vanderlocht
Mark A Kirkland
Reinout Hesselink
Wilfred TV Germeraad
Gerard MJ Bos
Publication date: 01-12-2011
Publisher: BioMed Central
Published in: BMC Immunology / Issue 1/2011
Electronic ISSN: 1471-2172
DOI: https://doi.org/10.1186/1471-2172-12-17

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