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A Novel In Vitro Model of Canine Malignant Hemangioendothelioma
Author(s) -
Kühn D.,
Kohn B.,
Brunnberg L.,
Plendl J.
Publication year - 2005
Publication title -
anatomia, histologia, embryologia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.34
H-Index - 35
eISSN - 1439-0264
pISSN - 0340-2096
DOI - 10.1111/j.1439-0264.2005.00669_64.x
Subject(s) - in vitro , angiogenesis , agglutinin , ulex europaeus , pathology , vascular endothelial growth factor , angiosarcoma , malignancy , cancer research , medicine , biology , lectin , immunology , vegf receptors , biochemistry
and Aim: Canine malignant haemangioendothelioma is an aggressive neoplasia that affects mostly older dogs of large breeds with a strong predilection for the spleen, liver, heart and skin. The tumour originates in the vascular endothelium and consists of transformed cells forming large and leaky vessel‐like structures. Prognosis is poor because surgery and chemotherapy have limited success in prolonging survival times and increasing quality of patients. A new strategy to treat this malignancy could be anti‐angiogenic therapy based on the inhibition of proliferation, migration and three‐dimensional organization of transformed cells. In order to reduce animal experiments, in vitro ‐models are required to test the safety and efficacy of anti‐angiogenic drugs. So far only few models of angiogenesis are available using mostly human, rodent and bovine cells. Therefore, the aim of our study was to establish an in vitro model of canine haemangioendothelioma. Materials and Methods: Tumours were collected from dogs during surgery or immediately after euthanasia. Isolation of cells was done from different areas of the tumours and by enzymatic digestion of the tissue. Cells were incubated in culture media with and without endothelial growth factors. Cells were characterized by lectin histochemistry using Dolichos biflorus agglutinin, Ulex europaeus agglutinin and Bandeiraea simplicifolia agglutinin I. Moreover, RT‐PCR (polymerase chain reaction) was employed to investigate the expression of vascular endothelial growth factor (VEGF) and its endothelium‐specific receptors VEGF‐R1 and ‐R2. Results and Conclusions: Different populations of cells were isolated and cultured successfully from canine malignant haemangioendothelioma. Cells show characteristics of microvascular endothelial cells of an angiogenic phenotype, i.e. the formation of spheroids and tube‐like structures as well as strong labelling for Bandeiraea simplicifolia agglutinin I. Thus, morphological and glycohistochemical results confirm the vascular character of the cells isolated. RT‐PCR showed expression of VEGF. However, endothelium‐specific VEGF receptors were not expressed. Loss of typical receptors is common in cancer and may correlate with increased tumour dedifferentiation.