In Vitro Modelling of Osteogenesis Imperfecta with Patient-Derived Induced Mesenchymal Stem Cells

• Published in: International journal of molecular sciences 2024-03, Vol.25 (6), p.3417
• Main Authors: Claeys, Lauria, Zhytnik, Lidiia, Ventura, Laura, Wisse, Lisanne E, Eekhoff, Elisabeth M W, Pals, Gerard, Bravenboer, Nathalie, Heine, Vivi M, Micha, Dimitra
• Format: Article
• Language: English
• Subjects: bone; Bone surgery; Cell Differentiation; cell model; Collagen; Collagen - metabolism; Fibroblasts; Flow cytometry; Gene expression; Humans; induced mesenchymal stem cells; Induced Pluripotent Stem Cells; Kinases; Medical research; Medicine, Experimental; Mesenchymal Stem Cells - metabolism; Mutation; osteoblast; Osteogenesis - genetics; Osteogenesis Imperfecta; Osteogenesis Imperfecta - genetics; Osteogenesis Imperfecta - metabolism; Pathophysiology; Patients; Proteins; skeletal dysplasia; Skin; Stem cells; T cell receptors; Transcription factors
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms25063417

(1) Mesenchymal stem cells (MSCs) are a valuable cell model to study the bone pathology of Osteogenesis Imperfecta (OI), a rare genetic collagen-related disorder characterized by bone fragility and skeletal dysplasia. We aimed to generate a novel OI induced mesenchymal stem cell (iMSC) model from induced pluripotent stem cells (iPSCs) derived from human dermal fibroblasts. For the first time, OI iMSCs generation was based on an intermediate neural crest cell (iNCC) stage. (2) Skin fibroblasts from healthy individuals and OI patients were reprogrammed into iPSCs and subsequently differentiated into iMSCs via iNCCs. (3) Successful generation of iPSCs from acquired fibroblasts was confirmed with changes in cell morphology, expression of iPSC markers , , and and three germ-layer tests. Following differentiation into iNCCs, cells presented increased iNCC markers including , , and and decreased iPSC markers, shown to reach the iNCC stage. Induction into iMSCs was confirmed by the presence of , , and markers, low expression of the hematopoietic, and reduced expression of the iNCC markers. iMSCs were trilineage differentiation-competent, confirmed using molecular analyses and staining for cell-type-specific osteoblast, adipocyte, and chondrocyte markers. (4) In the current study, we have developed a multipotent in vitro iMSC model of OI patients and healthy controls able to differentiate into osteoblast-like cells.