Use of a Collagen–Elastin Matrix as Transport Carrier System to Transfer Proliferating Epidermal Cells to Human Dermis in Vitro

Cell Transplantation

Article

Susan Gibbs¹  Taco Waaijman¹  Melanie Breetveld¹  Rik J. Scheper²  Magda Ulrich³  Esther Middelkoop³ 

[1] Department of Dermatology, VU University Medical Centre, Amsterdam, The Netherlands;Department of Pathology, VU University Medical Centre, Amsterdam, The Netherlands;Plastic, Reconstructive & Hand Surgery, VU University Medical Centre, Amsterdam, The Netherlands; Association of Dutch Burn Centers, Beverwijk, The Netherlands;

关键词: Transfer;    Keratinocyte;    Melanocyte;    Burns;    Wound healing;   

DOI  :  10.3727/096368910X507196

 received in 2009-12-04, accepted in 2010-04-29,  发布年份 2010

来源: Sage Journals

【 摘 要 】

This in vitro study describes a novel cell culture, transport, and transfer protocol that may be highly suitable for delivering cultured proliferating keratinocytes and melanocytes to large open skin wounds (e.g., burns). We have taken into account previous limitations identified using other keratinocyte transfer techniques, such as regulatory issues, stability of keratinocytes during transport (single cell suspensions undergo terminal differentiation), ease of handling during application, and the degree of epidermal blistering resulting after transplantation (both related to transplanting keratinocyte sheets). Large numbers of proliferating epidermal cells (EC) (keratinocytes and melanocytes) were generated within 10–14 days and seeded onto a three-dimensional matrix composed of elastin and collagen types I, III, and V (Matriderm®), which enabled easy and stable transport of the EC for up to 24 h under ambient conditions. All culture conditions were in accordance with the regulations set by the Dutch Central Committee on Research Involving Human Subjects (CCMO). As an in vitro model system for clinical in vivo transfer, the EC were then transferred from Matriderm onto human acellular dermis during a period of 3 days. After transfer the EC maintained the ability to regenerate into a fully differentiated epidermis containing melanocytes on the human dermis. Proliferating keratinocytes were located in the basal layer and keratin-10 expression was located in differentiating suprabasal layers similar to that found in human epidermis. No blistering was observed (separation of the epidermis from the basement membrane). Keratin-6 expression was strongly upregulated in the regenerating epidermis similar to normal wound healing. In summary, we show that EC-Matriderm contains viable, metabolically active keratinocytes and melanocytes cultured in a manner that permits easy transportation and contains epidermal cells with the potential to form a pigmented reconstructed epidermis. This in vitro study has produced a robust protocol that is ready for clinical studies in the future.

【 授权许可】

Unknown   
© 2010 Cognizant Comm. Corp.

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