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Blaimauer, K; Watzinger, E; Erovic, BM; Martinek, H; Jagersberger, T; Thurnher, D.
Effects of epidermal growth factor and keratinocyte growth factor on the growth of oropharyngeal keratinocytes in coculture with autologous fibroblasts in a three-dimensional matrix.
Cells Tissues Organs. 2006; 182(2):98-105
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Autor/innen der Med Uni Graz:
Thurnher Dietmar
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Abstract:
Tissue engineering of oropharyngeal mucosa is rendered complex by the fact that oropharyngeal keratinocytes are difficult to culture in the long term and do not grow well after several subcultivations. Three populations of oropharyngeal keratinocytes were isolated by a method based on different levels of beta(1)-integrin expression. In particular, keratinocytes were isolated between cell fractions that adhere rapidly on collagen-IV-coated culture dishes (RAC-IV) and populations that are less adherent (RAC-IV-D). The total fraction of both subpopulations served as a control (RAC-IV-T). The epidermal growth factor (EGF) and the keratinocyte growth factor (KGF) were examined with regard to their effects on the growth of the three populations. Growth curves of all three cell fractions grown with or without EGF were generated, and different concentrations of EGF and KGF were tested. EGF did not change any growth characteristics of the cells, with the exception of the speed of growth. Best growth was achieved with a physiologic EGF concentration of 0.15-1.5 ng/ml and a KGF concentration of 15 ng/ml. Finally, we cocultured oropharyngeal keratinocytes and their autologous fibroblasts in a three-dimensional matrix using Matrigeltrade mark. Oropharyngeal keratinocytes grown in coculture formed larger colonies than keratinocytes grown without fibroblasts. In conclusion, we were able to optimize the supplement of EGF and KGF in standard medium for the long-term culture of primary oropharyngeal keratinocytes. The use of Matrigel as a scaffold for three-dimensional cocultures of oropharyngeal keratinocytes and fibroblasts might signify a step forward in the development of a transplantable mucosa construct. Copyright 2006 S. Karger AG, Basel.
Find related publications in this database (using NLM MeSH Indexing)
Biocompatible Materials - chemistry
Cell Differentiation - drug effects
Cell Proliferation - drug effects
Cells, Cultured -
Coculture Techniques - methods
Collagen - chemistry
Drug Combinations -
Epidermal Growth Factor - metabolism
Epidermal Growth Factor - pharmacology
Fibroblast Growth Factor 7 - metabolism
Fibroblast Growth Factor 7 - pharmacology
Fibroblasts - physiology
Keratinocytes - cytology
Keratinocytes - drug effects
Laminin - chemistry
Mouth Mucosa - cytology
Oropharynx - cytology
Oropharynx - drug effects
Proteoglycans - chemistry
Tissue Engineering - methods

Find related publications in this database (Keywords)
epidermal growth factor
keratinocyte growth factor
matrigel
coculture
oropharyngeal keratinocytes
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