Authors: K Jubin Y Martin D J LawrenceWatt J R Sharpe
Publish Date: 2011/08/06
Volume: 63, Issue: 6, Pages: 655-662
Abstract
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 singlecell 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 61 excess of keratinocytes to fibroblasts this coculture 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 coculture 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 coculture 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
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