caa a22 4500 445823054 CHVBK 20180317145250.0 cr unu---uuuuu 170323e20111201xx s 000 0 eng 10.1007/s10616-011-9382-5 doi (NATIONALLICENCE)springer-10.1007/s10616-011-9382-5 A fully autologous co-culture system utilising non-irradiated autologous fibroblasts to support the expansion of human keratinocytes for clinical use [Elektronische Daten] [K. Jubin, Y. Martin, D. Lawrence-Watt, J. Sharpe] Autologous keratinocytes can be used to augment cutaneous repair, such as in the treatment of severe burns and recalcitrant ulcers. Such cells can be delivered to the wound bed either as a confluent sheet of cells or in single-cell suspension. The standard method for expanding primary human keratinocytes in culture uses lethally irradiated mouse 3T3 fibroblasts as feeder cells to support keratinocyte attachment and growth. In an effort to eliminate xenobiotic cells from clinical culture protocols where keratinocytes are applied to patients, we investigated whether human autologous primary fibroblasts could be used to expand keratinocytes in culture. At a defined ratio of a 6:1 excess of keratinocytes to fibroblasts, this co-culture method displayed a population doubling rate comparable to culture with lethally irradiated 3T3 cells. Furthermore, morphological and molecular analysis showed that human keratinocytes expanded in co-culture with autologous human fibroblasts were positive for proliferation markers and negative for differentiation markers. Keratinocytes expanded by this method thus retain their proliferative phenotype, an important feature in enhancing rapid wound closure. We suggest that this novel co-culture method is therefore suitable for clinical use as it dispenses with the need for lethally irradiated 3T3 cells in the rapid expansion of autologous human keratinocytes. Springer Science+Business Media B.V., 2011 Cell culture nationallicence Tissue engineering nationallicence Skin nationallicence Keratinocyte nationallicence Jubin K. Blond McIndoe Research Foundation, Queen Victoria Hospital, Holtye Road, East Grinstead, RH19 3DZ, West Sussex, UK aut Martin Y. Blond McIndoe Research Foundation, Queen Victoria Hospital, Holtye Road, East Grinstead, RH19 3DZ, West Sussex, UK aut Lawrence-Watt D. Blond McIndoe Research Foundation, Queen Victoria Hospital, Holtye Road, East Grinstead, RH19 3DZ, West Sussex, UK aut Sharpe J. Blond McIndoe Research Foundation, Queen Victoria Hospital, Holtye Road, East Grinstead, RH19 3DZ, West Sussex, UK aut Cytotechnology Springer Netherlands 63/6(2011-12-01), 655-662 0920-9069 63:6<655 2011 63 10616 https://doi.org/10.1007/s10616-011-9382-5 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10616-011-9382-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Jubin K. Blond McIndoe Research Foundation, Queen Victoria Hospital, Holtye Road, East Grinstead, RH19 3DZ, West Sussex, UK aut NATIONALLICENCE 700 1- Martin Y. Blond McIndoe Research Foundation, Queen Victoria Hospital, Holtye Road, East Grinstead, RH19 3DZ, West Sussex, UK aut NATIONALLICENCE 700 1- Lawrence-Watt D. Blond McIndoe Research Foundation, Queen Victoria Hospital, Holtye Road, East Grinstead, RH19 3DZ, West Sussex, UK aut NATIONALLICENCE 700 1- Sharpe J. Blond McIndoe Research Foundation, Queen Victoria Hospital, Holtye Road, East Grinstead, RH19 3DZ, West Sussex, UK aut NATIONALLICENCE 773 0- Cytotechnology Springer Netherlands 63/6(2011-12-01), 655-662 0920-9069 63:6<655 2011 63 10616 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer