Abstract
Tissue microarray (TMA) technique is an established high-throughput method to analyze multiple tissue specimens in parallel. However, in order to obtain reliable results from immunohistochemical analyses of TMA blocks, cell composition of TMA spots must correspond to whole tissue sections (WTS) particularly in tissues with a heterogeneous cell composition as it is the case in myeloproliferative neoplasms (MPN). The aim of this study was to validate TMA of bone marrow biopsies from MPN patients. TMAs of MPN bone marrow biopsies (ET: n = 26, PV: n = 26, and PMF: n = 29) were compiled in triplicates and MPN-specific histological parameters were assessed. Results of TMA spots were compared with WTS’ results using the intra-class correlation coefficient (ICC). Immunohistochemical NFE2 and calreticulin stainings of the TMA with quantitative evaluation were performed. TMA construction was technically successful with a loss of 10 % of all spots. ICC calculation revealed high to moderate correlations of TMA with WTS, especially the parameters that are typically affected in MPN tissue, e.g. cellularity of hematopoiesis (ICC 0.62–0.89), number of megakaryocytes (ICC 0.50–0.71), micro-vessel density (ICC 0.56–0.91), or grade of myelofibrosis (ICC 0.56–0.89). Results of NFE2 and calreticulin immunohistochemistry of MPN TMAs are consistent with previously published data. Overall, our results show moderate to good correlation between histological data of WTS and TMA spots illustrating that the TMA technique is applicable to bone marrow biopsies of MPN patients. However, TMA construction in triplicates is necessary to reach sufficient correlation. MPN TMAs can be applied for serial immunohistochemical surveys of archived tissues to assess the mutation status or to further sub-classify MPN cases.
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Acknowledgments
The authors thank Jasmin Roth, Markus Kühs, and Evelyn Wätzig for best technical support.
Author contribution
KA, MW, HLP, and DH planed the experiments, KL, RT, BH, and KA conducted the experiments and gathered data, KL, KA, LB, and DH analyzed data, and all authors wrote the manuscript.
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All Authors declare that there are no financial and non-financial competing interests.
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The study was approved by the local Ethics Committee, University of Freiburg (324/09).
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Limberger, K.A., Bogatyreva, L., Todorova, R. et al. Tissue microarray technique is applicable to bone marrow biopsies of myeloproliferative neoplasms. Histochem Cell Biol 147, 75–82 (2017). https://doi.org/10.1007/s00418-016-1476-x
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DOI: https://doi.org/10.1007/s00418-016-1476-x