The Potentials and Pitfalls of a Human Cervical Organoid Model Including Langerhans Cells

Abstract

Three-dimensional cell culturing to capture a life-like experimental environment has become a versatile tool for basic and clinical research. Mucosal and skin tissues can be grown as “organoids” in a petri dish and serve a wide variety of research questions. Here, we report our experience with human cervical organoids which could also include an immune component, e.g., Langerhans cells. We employ commercially available human cervical keratinocytes and fibroblasts as well as a myeloid cell line matured and purified into langerin-positive Langerhans cells. These are then seeded on a layer of keratinocytes with underlying dermal equivalent. Using about 10-fold more than the reported number in healthy cervical tissue (1–3%), we obtain differentiated cervical epithelium after 14 days with ~1% being Langerhans cells. We provide a detailed protocol for interested researchers to apply the described “aseptic” organoid model for all sorts of investigations—with or without Langerhans cells.

Characterization of 3D organotypic epithelial tissues reveals tonsil-specific differences in tonic interferon signaling

“…While modifications can be made to these 3D models depending on the experimental context and complexity desired, we used an organotypic 3D raft model with the same fibroblasts, collagen, media, growth conditions, and duration to maximize our ability to compare between tissues. For example, 3D organotypic skin and cervical models have been embedded with additional cell types, such as Langerhans cells (Jackson et al, 2020a;Kosten et al, 2015), and others have tested tissue-specific hormone supplementation (estrogen and progesterone) when culturing 3D organotypic ectocervical epithelium (McKinnon et al, 2020). A primary goal of our study was to provide evidence that these different in vitro tissues maintain similarities with their in vivo tissue equivalents, independent of other cell types or microbiota normally present in these tissues.…”

Characterization of 3D organotypic epithelial tissues reveals tonsil-specific differences in tonic interferon signaling

“…While modifications can be made to these 3D models depending on the experimental context and complexity desired, we used an organotypic 3D raft model with the same fibroblasts, collagen, media, growth conditions, and duration to maximize our ability to compare between tissues. For example, 3D organotypic skin and cervical models have been embedded with additional cell types, such as Langerhans cells [109,110], and others have tested tissue-specific hormone supplementation (estrogen and progesterone) when culturing 3D organotypic ectocervical epithelium [100]. A primary goal of our study was to provide evidence that these different in vitro tissues maintain similarities with their in vivo tissue equivalents, independent of other cell types or microbiota normally present in these tissues.…”