Vascular endothelial growth factor-A165 (VEGF-A165) and fibroblast growth factor-2 (FGF-2) are currently used for the functionalization of biomaterials designed for tissue engineering. We have developed a new simple method for heterologous expression and purification of VEGF-A165 and FGF-2 in the yeast expression system of Pichia pastoris. The biological activity of the growth factors was assessed in cultures of human and porcine adipose tissue-derived stem cells (ADSCs) and human umbilical vein endothelial cells (HUVECs). When added into the culture medium, VEGF-A165 stimulated proliferation only in HUVECs, while FGF-2 stimulated the proliferation of both cell types. A similar effect was achieved when the growth factors were pre-adsorbed to polystyrene wells. The effect of our recombinant growth factors was slightly lower than that of commercially available factors, which was attributed to the presence of some impurities. The stimulatory effect of the VEGF-A165 on cell adhesion was rather weak, especially in ADSCs. FGF-2 was a potent stimulator of the adhesion of ADSCs but had no to negative effect on the adhesion of HUVECs. In sum, FGF-2 and VEGF-A165 have diverse effects on the behavior of different cell types, which maybe utilized in tissue engineering.

Department of Biochemistry University of Chemistry and Technology Prague Technická 6 CZ 16628 Praha 6 Czech Republic

Department of Physiology Faculty of Science Charles University Viničná 7 CZ 12844 Praha 2 Czech Republic

Faculty of Biomedical Engineering Czech Technical University Prague CZ 27201 Kladno Czech Republic

Laboratory of Biomaterials and Tissue Engineering Institute of Physiology of the Czech Academy of Sciences Vídeňská 1083 CZ 14220 Praha 4 Czech Republic

Laboratory of Biotransformation Institute of Microbiology of the Czech Academy of Sciences Vídeňská 1083 CZ 14220 Praha 4 Czech Republic

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