Annals of Hematology

, Volume 92, Issue 5, pp 595–604

Cytological characterization of murine bone marrow and spleen hematopoietic compartments for improved assessment of toxicity in preclinical gene marking models

  • Min Yang
  • Guntram Büsche
  • Arnold Ganser
  • Zhixiong Li
Original Article


Gene therapy has proven its potential to cure diseases of the hematopoietic system, but potential adverse reactions related to insertional mutagenesis by integrating gene vectors and chromosomal instability in long-lived repopulating cells have emerged as a major limitation. Preclinical gene therapy in murine models is a powerful model for assessment of gene marking efficiency and adverse reactions. However, changes in the hematologic composition after transplantation with retrovirally modified hematopoietic stem cells have not been well investigated in large cohorts of animals by systematic cytological analyses. In the present study, cytological analyses of bone marrow and spleen were performed in a large cohort (n = 58) of C57BL/6J mice over an extended observation period after gene marking. Interestingly, we observed hematological malignancies in four out of 30 animals transplanted with dLNGFR (truncated form of the human p75 low-affinity nerve growth factor receptor) and tCD34 modified stem/progenitor cells. Our data demonstrate that cytological analysis provides important information for diagnosis of hematological disorders and thus should be included in preclinical studies and performed in each investigated animal. Together with histological analysis, flow cytometric analysis, and other analyses, the quality and predictive value of preclinical gene therapy studies will be improved.


Preclinical gene therapy studies Cytological analysis dLNGFR tCD34 

Supplementary material

277_2012_1655_MOESM1_ESM.doc (130 kb)
ESM 1(DOC 130 kb)
277_2012_1655_MOESM2_ESM.pdf (50 kb)
ESM 2(PDF 50.0 kb)
277_2012_1655_MOESM3_ESM.pdf (55 kb)
ESM 3(PDF 54.9 kb)
277_2012_1655_MOESM4_ESM.pptx (99 kb)
ESM 4(PPTX 98.9 kb)

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Min Yang
    • 1
  • Guntram Büsche
    • 2
  • Arnold Ganser
    • 3
  • Zhixiong Li
    • 1
  1. 1.Institute of Experimental Hematology, OE6960Hannover Medical SchoolHannoverGermany
  2. 2.Institute of PathologyHannover Medical SchoolHannoverGermany
  3. 3.Department of Hematology, Hemostasis, Oncology, and Stem Cell TransplantationHannover Medical SchoolHannoverGermany

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