Automated production of cultured epidermal autografts and sub-confluent epidermal autografts in a computer controlled bioreactor

• Published in: Biotechnology and bioengineering 1998-09, Vol.59 (6), p.679-683
• Main Authors: Prenosil, J.E., Villeneuve, P.E.
• Format: Article
• Language: English
• Subjects: Animal cells; Biological and medical sciences; bioreactor; Bioreactors; Biotechnology; Carbon Dioxide - metabolism; Cell Division; Culture Techniques - instrumentation; Culture Techniques - methods; cultured epidermal autografts; Epidermis - metabolism; Establishment of new cell lines, improvement of cultural methods, mass cultures; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; Humans; Hydrogen-Ion Concentration; keratinocyte cultures; Keratinocytes - cytology; Keratinocytes - metabolism; Kinetics; Methods. Procedures. Technologies; Software; Temperature; Time Factors; tissue engineering; Transplantation, Autologous - instrumentation; Transplantation, Autologous - methods
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/(SICI)1097-0290(19980920)59:6<679::AID-BIT4>3.0.CO;2-C

The objective of this work was to engineer an automated system for the production of cultured epidermal autografts and sub‐confluent cultured epidermal autografts. Human epidermal cells were grown directly on a transparent FEP film, which was held in place and surrounded by a polycarbonate growth chamber. The growth chambers were stacked to accommodate various surface area requirements. To monitor the development of the grafts, the upper‐most growth chamber in the stack was periodically placed on a standard phase contrast microscope. The growth chambers were connected to a multi‐channel peristaltic pump, which was controlled automatically to manage fluid‐handling operations. Sub‐confluent graft production involved removing the epidermal‐film composite from the growth chambers and cutting desired graft geometries. Producing cultured epidermal autografts involved (1) removing the confluent epidermal‐film composite from the growth chambers, (2) treating the composites with dispase, and (3) clipping the detached cultured epidermis to a synthetic support. Twelve to fifteen days were required to produce sub‐confluent grafts (total surface area 3500–4500 cm2 50% confluent) and 18 to 24 d were required to produce standard cultured epidermal autografts (total surface area 3500–4500 cm2). The system reduces the tedious manual labor associated with producing cultured epidermal autografts. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:679–683, 1998.