Establishing a transport protocol for the delivery of melanocytes and keratinocytes for the treatment of vitiligo.
We have previously developed a cell delivery and transfer technology for delivering autologous keratinocytes and melanocytes to patients with vitiligo. However, for this technology to benefit many patients geographically distant from the cell culture facility transportation issues need to be overcome. In this study we begin to investigate this by looking at what role surface chemistry and medium supplements, including fetal calf serum, CO₂ gassing, and temperature, play in influencing cell viability. Cells were maintained on carriers for up to 48 h outside of a CO₂ incubator at 37 °C and their subsequent ability to adhere and become organized into a new epithelium with appropriately located melanocytes was assessed. Consistently good viability and performance on an in vitro wound bed model was achieved by maintaining cells for 48 h adherent to a 20% acrylic acid coated carrier at lower (around 23 °C rather than 37 °C) temperatures in the medium preperfused with CO₂ before transport. Under these circumstances fetal calf serum was not required. In summary, the surface chemistry of the transport substrate and an appropriately CO₂ buffered medium at near room temperature can extend the effective performance life of these cultured cells to at least 48 h from when they leave standard incubator conditions.
Department of Engineering Materials, The Kroto Research Institute, University of Sheffield, Sheffield, United Kingdom.
Paula C
PC
Eves
Marta
M
Baran
Nial A
NA
Bullett
Louise
L
Way
David
D
Haddow
Sheila
S
Mac Neil
D001692:Biological Transport / Q000187:drug effects; D002245:Carbon Dioxide / Q000494:pharmacology; D002452:Cell Count; D002470:Cell Survival / Q000187:drug effects; D017690:Cell Transplantation / Q000379:methods*; D002478:Cells, Cultured; D018920:Coculture Techniques; D003470:Culture Media / Q000494:pharmacology; D020405:Dermis / Q000166:cytology / Q000187:drug effects; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects; D008544:Melanocytes / Q000166:cytology* / Q000187:drug effects; D013696:Temperature; D014820:Vitiligo / Q000628:therapy*
D003470:Culture Media; D002245:Carbon Dioxide