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Int. J. Mol. Sci. 2013, 14(3), 4684-4704; doi:10.3390/ijms14034684
Article

Irradiated Human Dermal Fibroblasts Are as Efficient as Mouse Fibroblasts as a Feeder Layer to Improve Human Epidermal Cell Culture Lifespan

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Received: 19 December 2012; in revised form: 7 February 2013 / Accepted: 19 February 2013 / Published: 26 February 2013
(This article belongs to the Special Issue Molecular Research of Epidermal Stem Cells)
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Abstract: A fibroblast feeder layer is currently the best option for large scale expansion of autologous skin keratinocytes that are to be used for the treatment of severely burned patients. In a clinical context, using a human rather than a mouse feeder layer is desirable to reduce the risk of introducing animal antigens and unknown viruses. This study was designed to evaluate if irradiated human fibroblasts can be used in keratinocyte cultures without affecting their morphological and physiological properties. Keratinocytes were grown either with or without a feeder layer in serum-containing medium. Our results showed that keratinocytes grown either on an irradiated human feeder layer or irradiated 3T3 cells (i3T3) can be cultured for a comparable number of passages. The average epithelial cell size and morphology were also similar. On the other hand, keratinocytes grown without a feeder layer showed heavily bloated cells at early passages and stop proliferating after only a few passages. On the molecular aspect, the expression level of the transcription factor Sp1, a useful marker of keratinocytes lifespan, was maintained and stabilized for a high number of passages in keratinocytes grown with feeder layers whereas Sp1 expression dropped quickly without a feeder layer. Furthermore, gene profiling on microarrays identified potential target genes whose expression is differentially regulated in the absence or presence of an i3T3 feeder layer and which may contribute at preserving the growth characteristics of these cells. Irradiated human dermal fibroblasts therefore provide a good human feeder layer for an effective expansion of keratinocytes in vitro that are to be used for clinical purposes.
Keywords: feeder layer; i3T3; Sp1; human; skin; keratinocyte; differentiation; proliferation; microarray; gene profiling feeder layer; i3T3; Sp1; human; skin; keratinocyte; differentiation; proliferation; microarray; gene profiling
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Bisson, F.; Rochefort, É.; Lavoie, A.; Larouche, D.; Zaniolo, K.; Simard-Bisson, C.; Damour, O.; Auger, F.A.; Guérin, S.L.; Germain, L. Irradiated Human Dermal Fibroblasts Are as Efficient as Mouse Fibroblasts as a Feeder Layer to Improve Human Epidermal Cell Culture Lifespan. Int. J. Mol. Sci. 2013, 14, 4684-4704.

AMA Style

Bisson F, Rochefort É, Lavoie A, Larouche D, Zaniolo K, Simard-Bisson C, Damour O, Auger FA, Guérin SL, Germain L. Irradiated Human Dermal Fibroblasts Are as Efficient as Mouse Fibroblasts as a Feeder Layer to Improve Human Epidermal Cell Culture Lifespan. International Journal of Molecular Sciences. 2013; 14(3):4684-4704.

Chicago/Turabian Style

Bisson, Francis; Rochefort, Éloise; Lavoie, Amélie; Larouche, Danielle; Zaniolo, Karine; Simard-Bisson, Carolyne; Damour, Odile; Auger, François A.; Guérin, Sylvain L.; Germain, Lucie. 2013. "Irradiated Human Dermal Fibroblasts Are as Efficient as Mouse Fibroblasts as a Feeder Layer to Improve Human Epidermal Cell Culture Lifespan." Int. J. Mol. Sci. 14, no. 3: 4684-4704.



Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert