Fluorescence-based tracing of transplanted intestinal epithelial cells using confocal laser endomicroscopy
Author(s)Bergenheim, Fredrik; Seidelin, Jakob B; Pedersen, Marianne T; Mead, Benjamin Elliott; Jensen, Kim B; Karp, Jeffrey Michael; Nielsen, Ole H; ... Show more Show less
MetadataShow full item record
BACKGROUND: Intestinal stem cell transplantation has been shown to promote mucosal healing and to engender fully functional epithelium in experimental colitis. Hence, stem cell therapies may provide an innovative approach to accomplish mucosal healing in patients with debilitating conditions such as inflammatory bowel disease. However, an approach to label and trace transplanted cells, in order to assess engraftment efficiency and to monitor wound healing, is a key hurdle to overcome prior to initiating human studies. Genetic engineering is commonly employed in animal studies, but may be problematic in humans due to potential off-target and long-term adverse effects. METHODS: We investigated the applicability of a panel of fluorescent dyes and nanoparticles to label intestinal organoids for visualization using the clinically approved imaging modality, confocal laser endomicroscopy (CLE). Staining homogeneity, durability, cell viability, differentiation capacity, and organoid forming efficiency were evaluated, together with visualization of labeled organoids in vitro and ex vivo using CLE. RESULTS: 5-Chloromethylfluorescein diacetate (CMFDA) proved to be suitable as it efficiently stained all organoids without transfer to unstained organoids in co-cultures. No noticeable adverse effects on viability, organoid growth, or stem cell differentiation capacity were observed, although single-cell reseeding revealed a dose-dependent reduction in organoid forming efficiency. Labeled organoids were easily identified in vitro using CLE for a duration of at least 3 days and could additionally be detected ex vivo following transplantation into murine experimental colitis. CONCLUSIONS: It is highly feasible to use fluorescent dye-based labeling in combination with CLE to trace intestinal organoids following transplantation to confirm implantation at the intestinal target site.
DepartmentDavid H. Koch Institute for Integrative Cancer Research at MIT; Massachusetts Institute of Technology. Institute for Medical Engineering and Science; Harvard University--MIT Division of Health Sciences and Technology
Stem Cell Research & Therapy
Bergenheim, Fredrik et al. "Fluorescence-based tracing of transplanted intestinal epithelial cells using confocal laser endomicroscopy." Stem Cell Research & Therapy 10 (May 2019): 148 DOI 10.1186/s13287-019-1246-5 ©2019 Author(s)
Final published version