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| Poster #265 Human olfactory organoids as an in vitro model of the olfactory epithelium |
| Kang-Hoon Kim1, Ankit Chauhan2, Noam Cohen1,2,3, Danielle Reed1, Peihua Jiang1, Hong Wang1, Jennifer E Douglas1,2 1Monell Chemical Senses Center, Philadelphia, PA, United States 2University of Pennsylvania, Department of Otorhinolaryngology - Head & Neck Surgery, Philadelphia, PA, United States 3Philadelphia Veterans Affairs Medical Center Surgical Services, Philadelphia, PA, United States |
The olfactory epithelium (OE) is a distinctive neuroepithelium capable of regeneration throughout life. However, injuries can lead to olfactory dysfunction, significantly affecting safety, nutrition, and quality of life. Direct studies of OE biology in humans have been limited due to restricted access to human tissue and lack of a human-based model system. We have successfully obtained human superior turbinate biopsies from adult and pediatric subjects, with tissue positive for olfactory cell markers on immunofluorescence, indicating the presence of OE. When cultured in media supplemented with niche factors, dissociated cells generate olfactory organoids. We performed immunostaining of the organoids using antibodies against OE markers, such as NCAM (immature and mature olfactory sensory neurons (OSNs)), olfactory marker protein (OMP; mature OSNs), SOX2 (globose basal cells (GBCs) and sustentacular cells (SCs)), keratin-5 (K5) (horizontal basal cells (HBCs)), and keratin-8 (K8) (SCs). Results demonstrate the presence of OE cell-type markers: NCAM+ cells have neuron-like morphology with a long process and a dendritic knob, cell body, and long axon, typical components of an OSN. We then performed RT-qPCR to quantify gene expression of OE cell types, demonstrating expression of markers for mature OSNs (OMP), SCs (SOX2, EZRIN), GBCs (SOX2), and HBCs (KRT5, SOX2). Organoids show Ca2+ release in response to odorant exposure, a surrogate for function. As a control, organoids cultured from inferior turbinate (non-OE) tissue do not mobilize Ca2+. This suggests organoids contain functional OSNs, further supporting the model. In summary, preliminary data supports a novel human olfactory organoid model which may help investigate OE biology in normal and diseased states, with potential for therapeutic insights. |