The proposed Bone Cells Analysis Core (BCAC) will enable participating investigators to pursue new lines of research in skeletal biology by using state-of-the-art technologies for bone cell isolation, functional assessment, and RNA sequencing. The proposed Core will bring together the expertise of its leadership (Director Dr. Pajevic Divieti and the Associate Directors Drs. Gardella, Gerstenfeld and Warman) and powerful new research approaches and assay systems to make for a synergistic Core Resource that will greatly enrich and catalyze the scientific advances of participating investigators. The BCAC will provide Center investigators (CIs) with 1) Preclinical and clinical bone cells and genetic tools, 2) Cell Signaling and Biomarker Assay services, and 3) RNA sequencing services. The Bone cell Core (Drs Divieti Pajevic and Gerstenfeld, Boston University), will provide mouse- derived bone cells and cells lines from both wild type or genetically modified animals. The Core will also provide, in collaboration with Dr. Gerstenfeld (Associate Director), highly annotated human specimens from total hip replacements. The BCAC will offer a wide array of in-house and commercially available signaling assays under the Signaling Assay service (Dr. Gardella, Associate Director, Endocrine Unit, MGH). The Core will help with the study design and will train investigators in all the aspects of these techniques. The RNA sequencing core (Dr. Warman, Associate Director, Boston Children?s Hospital) will provide services to help CIs and their trainees design, perform, and analyze bulk and single cell RNA sequencing experiments that involve skeletal cells and tissues. The Core will provide help with designing bulk and single cell RNA sequencing experiments and with assisting with tissue and cell preparation for RNA recovery. The core will provide micro-pulverizers and optimized methods for recovering RNA from bone and cartilage for bulk RNA sequencing and living cells for single cell RNA sequencing. The Core will generate sequencing libraries using kits designed for very low amounts of starting RNA. The Core will assist in the informatics analysis of the sequencing data. Each service provided by the Core will be rigorously controlled for quality performance and reliability. The Core will conduct hands-on training sessions by which investigators can be mentored and taught how to perform assays, cell manipulations on their own and RNA sequencing. It will also establish a seminar series in which new methodologies, approaches, and related scientific progress will be presented and discussed. !
