Project Details
Projekt
Generation of human induced pluripotent stem cells
Applicant
Professor Dr. Arne Hansen
Subject Area
Orthopaedics, Traumatology, Reconstructive Surgery
Term
since 2024
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 517063424
In the context of the clinical research unit ProBone (“Precision Medicine for Early-Onset Low Bone Mineral Density Disorders”) this central project (CP1) will provide human induced pluripotent stem cell (hiPSC) models for molecular studies of gene variants causing an early-onset low bone mineral density (BMD) disorder. Since hiPSCs have evolved as an important human in vitro model for biomedical research in the past years, CP1 will enable an important extension of the precision medicine approach applied in the different ProBone projects. In the first work package CP1 will establish a biobank of primary urinary cells (PUCs) from early-onset low BMD patients. PUCs from specific patients relevant for the respective ProBone projects will be reprogrammed to hiPSCs by sendai virus-mediated delivery of reprogramming factors. In the second work package, CP1 will optimize and scale up protocols to differentiate osteoblasts and establish respective protocols for chondroblasts and osteoclasts as relevant osteogenic target cell types to study disease mechanisms of specific early-onset low BMD disorders. As complementary 3D assay formats CP1 will establish bone micro-physiological and organoid models based on published protocols and future progress in the field. In the third work package low BMD disorder-defining gene variants in the patient-derived hiPSCs lines will be corrected by CRISPR/Cas9 genetic engineering to establish cell line-specific isogenic controls. Basis for these work packages is a well-established standard operating procedure-based platform for hiPSC- and CRIPSR/Cas9-technologies in the Institute for Experimental Toxicology and Pharmacology. The isogenic hiPSCs lines, differentiation protocols for osteogenic cell types and 3D models will be shared with and transferred to the respective ProBone projects to advance understanding of disease mechanisms and to develop targeted treatment concepts. Overall, this central project will establish a human in vitro platform to study early-onset low BMD disorders as a complementary approach to existing animal models.
DFG Programme
Clinical Research Units
