Abstract

University of Connecticut School of Medicine and Dentistry and Scripps Research Institute  TMC: High resolution 3D mapping of cellular heterogeneity within articular cartilage

This recently expanded group will utilize a protocol that is capable of performing multimodal histology that includes methods for repetitive imaging of in situ hybridization and antibody binding. This will be applied to the distal end of the femur with a primary focus on the articular cartilage.  

The team acquires femoral tissues from de-identified human sources who have not had identifiable articular cartilage abnormalities. Each cartilage and meniscus  sample is oriented to its source tissue and defined section utilized for multiple analytical workflows to identify the molecular fingerprint and location of each cell with the sample.  The analysis include scRNAseq  liberated intact chondrocytes and chondrocyte nuclei, spatial distribution of the RNA fingerprints (Visium and 10X Genomics)  and ultimately direct cellular location of specific cell types by both in situ RNA hybridization (MERFISH) and proteins (CODEX).

This team will also provide opportunities to transfer the histological technology to major academic skeletal research groups. Once the skeletal data becomes available, they will develop virtual workshops to inform the skeletal biology community of this valuable resource and how it can be utilized to unravel rare diseases affecting the skeleton. HuBMAP will be a transformational technology that every tissue centric group needs to incorporate. This team’s role is to ensure that the skeletal biology community is included in this new experimental platform, and that it is employed to solve the major genetic and therapeutic challenges affecting skeletal health.