Establishing a Transport Protocol for the Delivery of Melanocytes and Keratinocytes for the Treatment of Vitiligo

Abstract

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.

Optimized Cell Survival and Seeding Efficiency for Craniofacial Tissue Engineering Using Clinical Stem Cell Therapy

“…Although temperature did not affect cell-seeding efficiency in our study, it is well established that temperature can have a profound effect on cell viability. In various tissue-grafting and organ-transplantation protocols, it is often highly desirable to maintain tissue specimens at 4°C (on ice) until ready for application or placement in the recipient site [31][32][33][34]. In regenerative cell-transplantation strategies involving stem cells, although important, this parameter has not been thoroughly examined.…”