title: Enrichment of unlabelled human Langerhans cells by modified discontinuous Ficoll-metrizoate density medium and following Langerhans cell culture.
abstract: Highly enriched and specifically unlabelled human Langerhans cells (LC) from epidermal cell suspensions (ECS) are indispensable for the intensive study of LC function. Discontinuous Ficoll-metrizoate density gradient centrifugation was found to be a satisfactory method of enriching LC from ECS. In contrast to a previous study, however, the majority of LC floated on Ficoll-metrizoate with a density of 1.057 g/cm3 instead of 1.068 g/cm3. The concentration of unlabelled LC can be enriched to as high as 92% from the original 1.8-3.1% LC in ECS. Approximately 40-50% of original LC can be harvested. The procedure was also simplified by using a Schick razor instead of a dermatome to obtain thin epidermal sheets (about 0.25 mm in thickness), omitting the density gradients of 1.089 and 1.10 g/cm3 and using the avidin-biotin complex method to identify LC. LC are non-proliferative cells in in vitro culture systems. The viability of LC dropped to 20-30% after 3 days in the culture medium. It was also observed that LC tended to attach to aggregated keratinocytes in the culture system.
authors.affiliation: Department of Dermatology, National Yang-Ming Medical College, Taiwan, R.O.C.
authors.forename: H N
authors.initials: HN
authors.lastname: Liu
authors.forename: Y T
authors.initials: YT
authors.lastname: Chang
authors.forename: C K
authors.initials: CK
authors.lastname: Wong
mesh_terms: D002470:Cell Survival; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D005362:Ficoll / Q000494:pharmacology; D016591:Histocytological Preparation Techniques; D006801:Humans; D007801:Langerhans Cells; D008793:Metrizamide / Q000494:pharmacology
chemical_list: D005362:Ficoll; D008793:Metrizamide