gms | German Medical Science

27th German Cancer Congress Berlin 2006

German Cancer Society (Frankfurt/M.)

22. - 26.03.2006, Berlin

Similarities and differences of human healthy and malignant hematopoietic stem cells

Meeting Abstract

  • corresponding author presenting/speaker Heinrich Lannert - Medizinische Klinik der Universität Heidelberg, Deutschland
  • Thomas Able - Medizinische Klinik der Universität Heidelberg
  • Thomas Franz - Proteomics Core Facility, EMBL Heidelberg
  • Anja Bathke - Proteomics Core Facility, EMBL Heidelberg
  • Reiner Hofmann - Medizinische Klinik, Ludwigshafen
  • Angela Lenze - Medizinische Klinik der Universität Heidelberg
  • Kerstin Horsch - Medizinische Klinik der Universität Heidelberg
  • Volker Eckstein - Medizinische Klinik der Universität Heidelberg
  • Reiner Saffrich - Medizinische Klinik der Universität Heidelberg
  • Anthony Dick Ho - Medizinische Klinik der Universität Heidelberg

27. Deutscher Krebskongress. Berlin, 22.-26.03.2006. Düsseldorf, Köln: German Medical Science; 2006. DocPO109

The electronic version of this article is the complete one and can be found online at:

Published: March 20, 2006

© 2006 Lannert et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.



Aim: The tracking of stem cell aging, differentiation and deterioration by gene expression profiling and proteome analysis allows the comparison of different stages. The overall aim of a proteomic study is characterization of the complex network of cell regulation. We focused our investigations on different subsets of highly enriched CD34+ stem cells from different human origins: cord blood (CB), bone marrow (BM), and mobilized stem cells from peripheral blood (PBSC), as well as CD34+ leukemia cells, thus e.g. to identify pathways and new targets for leukemia therapy.

Methods: Mononuclear cells were isolated by a standard Ficoll-Hypaque gradient separation method from the different blood sources. An Auto-MACS (Miltenyi) and FACS Vantage SE cell sorter (Becton Dickinson) was used to highly enrich (>99%) CD34+ cells fractions. Sample preparation: Total RNA was isolated from sorted 1 x 10e6 cells by standard methods using RNA isolation kit (Qiagen). For gene expression analysis topic-defined PIQOR™ stem cell microarrays (936 genes) were performed. Proteomics started with the determination of protein concentrations, 2D-gel-electrophoresis were described in Proteome Works System (BioRad). Sypro ruby and/or coomasie stained gels were used for protein identification by Q-TOF analyses. The subcellular localization of the identified proteins were performed by fluorescence and confocal microscopy of all cell fractions.

Results: 1. The microarray gene expression correlation shows many similarities between human healthy stem cells of different sources and ages, otherwise many differences: 125 upregulated genes (e.g. kinases: PAK1, ATM1, CDKN2A) and 32 downregulated genes in malignant cells compared to healthy stem cells. 2. The protein pattern of the CD34+ highly enriched stem cell fractions is complex and shows >1000 protein spots (dependent on protein concentration and staining) using PDQuest software. 3. The most dominant protein spots (>100) were cutted and identified by Q-TOF. There is a strong overlap between the detected major proteins in all analyzed cell fractions. 4. E.g. stathmin (oncoprotein Op18) is expressed at very high levels in leukemia cells and in PBSCs but not in BM cells, additionally demonstrated by fluorescence microscopy.

Conclusions: Using Genomics and Proteomics assays, pathways (substrate, kinases) e.g. for proliferation and migration of healthy (mobilized) CD34+ cells from bone marrow and malignant leukemia cells could be identified.