![]() ![]() ![]() Basic fibroblast growth factor plays an important role in the proliferation and differentiation processes of a wide range of cells. This strategy is commonly applied to adsorb basic fibroblast growth factor (FGF-2) on heparinized surfaces. In some cases, it is desirable to attach the protein molecule to the surface by making use of its natural affinity to the specific biological substances. Covalent binding methods also show low selectivity, thus resulting in co-immobilization of the impurities present in the protein solution. Covalent immobilization can affect the stability and the bioactivity of biomolecules because of the random orientation of the bounded protein, or can lead to the loss of functional groups present on the protein surface due to coupling reactions. The surface can be modified with proteins by direct covalent attachment of the biomolecule to the support, or by physicochemical adsorption based on specific and/or non-specific interactions between the biomolecule and the surface. Properly designed substrates can stabilize the growth factor molecules, enhance activity and control their release. ![]() It has been shown that such systems can enhance cell growth due to optimized local concentration of the growth factor, which results in stronger interactions with the cell membrane receptors. Alternatively, direct immobilization of growth factors on the cell culture surfaces may provide a constant reservoir of bio-signaling molecules without the need for additional supplementation of the culture medium with growth factors. In a typical cell culture system, systematic administration of growth factors in the culture medium is required to stimulate cell growth. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have declared that no competing interests exist. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are creditedĭata Availability: All relevant data are within the paper and its Supporting Information files.įunding: The study was funded by the Ministry of Education, Youth and Sports, Czech Republic (grant number EE2.), by the Grant Agency of the Czech Republic (grant number P1 and grant number P108/12/P624), and by the European Regional Development Fund (Project BIOCEV: Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University, grant number CZ.1.05/1.1.00/02.0109). Received: DecemAccepted: MaPublished: May 6, 2015Ĭopyright: © 2015 Kumorek et al. PLoS ONE 10(5):Īcademic Editor: Jui-Yang Lai, Chang Gung University, TAIWAN (2015) Cellular Responses Modulated by FGF-2 Adsorbed on Albumin/Heparin Layer-by-Layer Assemblies. The improvement was more pronounced at a lower FGF-2 surface concentration (30 ng/cm 2) than on surfaces with a higher concentration of FGF-2 (120 ng/cm 2).Ĭitation: Kumorek M, Kubies D, Filová E, Houska M, Kasoju N, Mázl Chánová E, et al. The adsorbed FGF-2 was bioactive and stimulated both the proliferation and the differentiation of CPAE cells. CPAE cells could attach and proliferate on Alb/Hep surfaces. The bioactivity of the adsorbed FGF-2 was assessed in experiments in vitro, using calf pulmonary arterial endothelial cells (CPAE). The results revealed that FGF-2 binds to the Alb/Hep assembly in a dose and time-dependent manner up to the surface concentration of 120 ng/cm 2. The adsorption of FGF-2 on the cross-linked Alb/Hep was followed by SPR. The formation of the Alb/Hep assemblies was monitored by the surface plasmon resonance (SPR), the infrared multiinternal reflection spectroscopy (FTIR MIRS) and UV/VIS spectroscopy. An analysis of the surface morphology by atomic force microscopy showed that two Alb/Hep bilayers are required to cover the surface of substrate. The surfaces were constructed by layer-by-layer adsorption of polyelectrolytes albumin and heparin and were subsequently stabilized by covalent crosslinking with glutaraldehyde. In this paper, we report on the fabrication of albumin/heparin (Alb/Hep) assemblies for controlled binding of basic fibroblast growth factor (FGF-2). In a typical cell culture system, growth factors immobilized on the cell culture surfaces can serve as a reservoir of bio-signaling molecules, without the need to supplement them additionally into the culture medium. ![]()
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