Characterization and modeling of cell activation upon TGF-β1 stimulation in a 3-D culture system

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.

Name thesis:degree_name

M.S.

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Bioengineering

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2010

Author dc:creator

• Xu, Jing

Contributors dc:contributor

• Insana, Michael F.

• myofibroblasts
• transforming growth factor-beta1
• alpha-smooth muscle actin
• diffusion
• confocal microscopy imaging
• immunofluorescence staining

Statement dc:rights

• Copyright 2010 Jing Xu

Language dc:language

en

Handle dc:identifier

http://hdl.handle.net/2142/16963

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/16963