Herstellung und Validierung von Tissue Microarrays von Knochenmarkbiopsien

Today, the construction of tissue microarrays in order to investigate the expression of proteins and genes on large case series is an approved and accepted method. The applicability of the tissue microarray technique for analysis of bone marrow biopsies has not been tested before and up to now not been mentioned in literature. The aim of our work was the construction of bone marrow TMAs, the analysis of arising technical difficulties and the immunohistochemical analysis of 15 markers used on the TMAs. The technique has been validated in a direct manner by the comparison with the corresponding full tissue sections and indirectly by the comparison with data from international literature. In our study we include 148 cases of acute leukemia, which comprise 115 AML (according to the FAB classification: 10 M0, 12 M1, 30 M2, 5 M3, 29 M4, 15 M5, 1 M6, 2 M7 und 11 not classified) and 33 ALL (23 B- and 5 T-ALL, 5 not classified). Some cases were punched twice, the others three times each. After the construction of the tissue microarrays we performed a morphological analysis of the H&E-stained section and an immunohistochemical analysis of the expression of the following markers: CD3, CD10, CD15, CD20, CD34, CD61, CD68, CD79a , CD99, CD117, CD138, MPO, HbA1, Glycophorin and TdT. The results can be summarized as follows: The TMA technique is an adequate method for the evaluation of bone marrow biopsies. In morphological analysis of the H&E-stained TMAs 6 (4%) of the 148 cases (384 punches overall) were not evaluable. 14 of the 384 punches (4%) because of missing tumor cells and 28 (7%) because of missing tissue (table 23). The number of sample losses is comparable to data of studies about TMAs on different tissues. By the immunohistochemical analysis of 15 markers on the three TMAs we obtained a number of 2220 analyses (each consisting of 2 resp. 3 punches) and 5760 single analyses, of which 723 (13%) were not evaluable, 437 (8%) of these because of missing tumor cells and 286 (5%) because of missing tissue (table 23). For the direct comparison of TMA cases with conventional sections we chose 7 of the 15 markers (CD3, CD10, CD20, CD34, CD68, CD117 und TdT. It revealed a concordance of 100%, which correlates well data in international literature, where a concordance between 82,5 - 100% was reached on different tissues. The results of the immunohistochemical analysis on maximum 0,6mm2 sized cores correspond surprisingly in 100% to those of the full section. In our study we were able to show that the utilisation of the TMA technique on bone marrow biopsies of acute leukemias is a reliable and effective method. Future large case cohort studies could be performed especially while using a combination of the TMA technique with DNA arrays with substantial time and cost savings. Dispite minor material losses due to the application of the TMA technique these studies can result in a broader and improved information content.

Grantor dc:publisher

Publikationsserver der RWTH Aachen University

Year dc:date

2007

Author dc:creator

• Schönberg, Sharon

Contributors dc:contributor

• Dirnhofer, Stefan

• info:eu-repo/classification/ddc/610
• Akute Leukämie
• Knochenmarkbiopsie
• Validierung
• Medizin
• Tissue Microarray
• Immunhistochemie
• acute leukemia
• bone marrow biopsy
• validation
• IHC

Statement dc:rights

• info:eu-repo/semantics/openAccess

Language dc:language

ger

Identifier

https://publications.rwth-aachen.de/search?p=id:%22RWTH-CONV-123933%22

OAI identifier oai:identifier

oai:publications.rwth-aachen.de:62362