Allah, Nasar Um Min
(2018)
Effect of fibroblast and platelet-derived growth factors on co-culture of human gingival fibroblasts and umbilical vein endothelial cells.
Masters thesis, Universiti Sains Malaysia.
Abstract
Numerous types of single cells in in-vitro cultures have been studied in tissue
engineering, but the study on direct paracrine interactions between heterotypic cells
population is lacking. Co-culture approach establishes an excellent atmosphere to
study these interactions. The objective of this in-vitro experimental study was to
determine the effects of fibroblast and platelet-derived growth factor ((FGF-2 and
PDGF-BB) in a co-culture of human gingival fibroblasts (HGFs) and human umbilical
vein endothelial cells (HUVECs). To this end, the medium for the establishment of
monolayer and co-culture of these cells were first optimised. Thereafter, the optimal
concentrations of these growth factors were determined in a monolayer and then in a
co-culture medium by assessing the cell viability using MTT assay. Next, gene
expression analysis for fibroblast and angiogenic biomarkers was assessed using realtime
RT-PCR to study the stimulatory effect of these growth factors by using 6 wellplate
with transwell inserts. Afterwards, statistical analysis of the results was
performed using one-way ANOVA and Kruskal-Wallis test with p < 0.05 considered
statistically significant. Results of cell viability assay showed that the effect of FGF-2
on HGF was dose-dependent and was optimum at a concentration of 5 ng/ml (p
=0.001), while that of PDGF-BB on HUVEC was optimum at a concentration of 20
ng/ml (p =0.004). The stimulatory effect of FGF-2 and PDGF-BB on HGF and
HUVEC was supported by the real-time RT-PCR results which showed that there is a
significant upregulation (p < 0.05) of gene biomarkers in the treatment group of bothcells after three days of co-culture experiment, compared to control group. Therefore,
it is concluded that there is the possibility of a synergistic effect of these two growth
factors on a co-culture of HGF and HUVEC which were suggestive of a proangiogenic
activity.
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