Overview
Topic status: We're looking for students to study this topic.
The successful isolation and ex–vivo expansion of human skin–derived keratinocytes has resulted in the use of these cells for therapeutic applications. However, currently the use of cultured keratinocytes carries a real risk of introducing infectious agents since the culture systems employed require the presence of animal serum, semi–defined or purified proteins, and/or “feeder” cells. In particular, we hypothesize that the co–culture of fibroblast feeder cells with keratinocytes results in the production of proteins/peptides that are not produced when these cells are cultivated alone. Furthermore, we propose that identification of these proteins/peptides will provide critical information that will facilitate the development of a culture systems that supports ex–vivo expansion of keratinocytes in the absence of BOTH animal feeder cells and serum.
This project will adopt an advanced proteomic approach to identify the proteins/peptides produced as a result of the autocrine, juxtacrine and paracrine interactions that occur between keratinocytes and fibroblast feeder cells. We will then assess whether incorporation of these candidate protein/peptides into our current serum–free medium results in a formulation that supports expansion of keratinocytes ex–vivo in the absence of both serum and feeder cells.
Methods and techniques
The methods and techniques that will be developed in the course of this project are:
- protein characterization – affinity purification, mass spectrometry, Nterminal sequencing
- proteomeLab™ PF 2D Protein Fractionation System
- Matrix Assisted Laser Desorption Ionization Time–of–Flight Time–of–Flight (MALDI–TOF–TOF) mass spectrometer
- cell culture – human skin culture
- immunohistochemistry, western blot analysis
- real–time PCR.
- Study level
- PhD, Masters, Honours
- Supervisors
- QUT
- Organisational unit
Science and Engineering Faculty
- Research area
- Contact
- Please contact a supervisor for enquiries.
Professor Zee Upton
Dr Gary Shooter
Dr Kerry Manton
