Nonirradiated human fibroblasts and irradiated 3T3-J2 murine fibroblasts as a feeder layer for keratinocyte growth and differentiation in vitro on a fibrin substrate.
The standard method for producing graftable epithelia relies on the presence of a feeder layer of lethally irradiated 3T3-J2 murine fibroblasts (Rheinwald and Green technique). Here, we studied a new keratinocyte culture system, which envisages the utilization of nonirradiated human fibroblasts embedded into a fibrin substrate, in cultures destined for a future clinical application. We tested this culture system using keratinocytes grown on a fibrin gel precoated with 3T3-J2 murine fibroblasts as a control. In order to evaluate the new technology, we compared the clonogenic potential and the proliferative, differentiative and metabolic characteristics of keratinocytes cultured on the fibrin gel under the two culture conditions. The results demonstrated that the proposed technology did not impair the behavior of cultured keratinocytes and revealed that cells maintained their proliferative potential and phenotype under the experimental conditions. In particular, the demonstration of stem cell maintenance under the adopted culture conditions is very important for acute burn treatment with skin substitutes. This work is a first step in the evaluation of a new keratinocyte culture system, which has been studied in order to take advantage of an additional human cell population (i.e. nonirradiated, growing fibroblasts) for future transplantation purposes in acute and chronic wounds. Additional research will allow us to attain (1) the removal of murine cells in the initial phase of keratinocyte cultures, and (2) the removal of other potentially dangerous animal-derived materials from the entire culture system.
Panacchia
Laura
L
Tissue Engineering and Cutaneous Physiopathology Laboratory, Istituto Dermopatico dell'Immacolata-IRCCS, Rome, Italy.
Dellambra
Elena
E
Bondanza
Sergio
S
Paterna
Patrizia
P
Maurelli
Riccardo
R
Paionni
Emanuel
E
Guerra
Liliana
L
D016475:3T3 Cells / Q000166:cytology* / Q000502:physiology / Q000528:radiation effects; D000818:Animals; D001672:Biocompatible Materials; D002450:Cell Communication; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D005337:Fibrin; D015718:Fibrin Tissue Adhesive; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000502:physiology; D051379:Mice; D013234:Stem Cells / Q000166:cytology / Q000502:physiology; D023822:Tissue Engineering
D001672:Biocompatible Materials; D015718:Fibrin Tissue Adhesive; D005337:Fibrin