Cell culture and basic fibroblast growth factor expression in canine hemangiosarcoma

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Trochta, Oliver Adam Kamil

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University of Guelph

Abstract

Hemangiosarcoma is a relatively common tumour of middle-aged to old dogs. Little is known concerning the biology of this tumour. To initiate work in this area, studies were designed to: (1) culture hemangiosarcoma cells in the hopes of establishing cell lines to be used in future studies and (2) examine the expression of basic fibroblast growth factor (FGF-2) in hemangiosarcomas since this growth factor is involved in angiogenic proliferation in other tumour systems and may function in an autocrine or paracrine fashion in canine hemangiosarcoma. von Willebrand factor (vWF), an endothelial marker, was used to identify and monitor the in vitro growth of normal and malignant endothelial cells. Of the two growth media used only M131 supplemented with a proprietary microvascular growth supplement facilitated the short-term culture of vWF positive cells from canine adrenals and rarely HS tumours. Reverse transcriptase polymerase chain reaction primers based on the human FGF-2 cDNA amplified a predicted 390 bp cDNA from canine tissues which showed 93% sequence homology with the human sequence. The cDNA product used as a probe in Northern blots, was unable to detect FGF-2 expression in RNA from bovine, human and canine tissues. Southern blot analysis showed that the probe hybridized to bovine, human and canine cDNA amplification products. FGF-2 antigen was demonstrated in hemangiosarcoma cells by immunohistochemistry, These results demonstrate that malignant cells in hemangiosarcoma elaborate FGF-2 and FGF-2 can be detected in RNA extracted from dogs by species specific RT-PCR primers. The presence of FGF-2 may have an important role in the growth of canine hemangiosarcoma.

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Keywords

cell culture, basic fibroblast growth factor, expression, angiogenic proliferation, canine hemangiosarcoma

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