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Title:Characterization and modeling of cell activation upon TGF-β1 stimulation in a 3-D culture system
Author(s):Xu, Jing
Advisor(s):Insana, Michael F.
Department / Program:Bioengineering
Degree Granting Institution:University of Illinois at Urbana-Champaign
transforming growth factor-beta1
alpha-smooth muscle actin
confocal microscopy imaging
immunofluorescence staining
Abstract:In this work, we explored a thick in vitro 3-D cell culture model with layered structure and applied transforming growth factor-beta1 (TGF-β1) on the top of the 3-D culture to induce the conversion from normal fibroblasts to myofibroblasts and expression of a marker protein, alpha-smooth muscle actin (α-SMA). The cell response with TGF-β1 stimulation was captured with immunofluorescence staining (IF) and confocal microscopy imaging of α-SMA in each cell embedded layer. Our results revealed the spatial-temporal profiles of the conversion from normal fibroblasts to myofibroblasts in 3-D culture upon TGF-β1stimulation. The conversion starting time, speed and saturation plateau are closely related to TGF-β1 concentration. Based on these experimental results, we successfully developed a mathematical model, which incorporates diffusion of growth factors and cell activation responses to describe the dynamic cell response to growth factors across the 3-D culture depth and predict the spatial-temporal profile of the fibroblast conversion within the culture. In summary, we present here a handy experimental and computational tool for researches on tumor signaling pathways, and provide some guidance for studies on diffusion of protein factors or drugs in 3-D tissue environment.
Issue Date:2010-08-31
Rights Information:Copyright 2010 Jing Xu
Date Available in IDEALS:2010-08-31
Date Deposited:2010-08

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