Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1233589 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR32621700_1.fastq.gz | Fastqsanger.gz | |||
| SRR32621700_2.fastq.gz | Fastqsanger.gz | |||
| SRR32621701_1.fastq.gz | Fastqsanger.gz | |||
| SRR32621701_2.fastq.gz | Fastqsanger.gz | |||
| SRR32621702_1.fastq.gz | Fastqsanger.gz |
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Project description:Study on gene expression during bone defect repair and the effects of heparin on bone tissue.
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Project description:Background: Mesenchymal stromal cells (MSCs) are a promising cell source for tissue engineering and regenerative medicine. In our lab, we found that cell preparations from bone marrow of many donors had a limited capacity for in vitro-differentiation into the osteogenic and chondrogenic lineages although this is a capacity claimed to be inherent to this kind of cells. Therefore the current study was designed to test the hypothesis whether the amount of heparin used as anti-coagulant during bone marrow harvest has inhibitory influence on the in vitro-differentiation capacity of the isolated MSCs. Methods: Bone marrow was obtained from twelve donors with good physical condition during preparation of the femoral cavity for the implantation of the femoral stem for a total hip arthroplasty in the absence or presence of heparin. Isolated MSCs were characterized by plastic adhesion, morphology, population doubling times, expression of cell surface antigens and in vitro-differentiation into the adipogenic, osteogenic and chondrogenic lineages. In addition, transcriptome analyses were performed. Results: Bone marrow prepared in the absence of heparin showed a comparable processability as bone marrow in the presence of heparin. Notably, no coagulation was observed. The number of mononuclear cells retrieved from the density gradient was independent of heparin addition. Moreover, none of the phenotypic or functional parameters assessed correlated with the addition of heparin. Transcriptome and quantitative realtime PCR analyses demonstrated that some selected genes were up- or downregulated in the cells isolated with the addition of heparin. However, this statement held true for only some but not all of the donors investigated in the present study. Conclusions: No correlation with heparin addition was observed with respect to the parameters assessed in the present study. This implies the absence of consequences both for the results of in vitro-investigations and also during in vivo-application, thereby ruling out heparin as a potential source of disparate results that are described in the literature.
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Project description:Temporal Gene Expression Profiling of Bone Repair in a Rat Calvarial Defect
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Project description:The vascular wall from diverse human organs is a source of mesenchymal progenitor cells. FACS purified human perivascular stem cells (PSC), identified by expression of CD146 or CD34, were observed to induce mitogenic, pro-migratory, and pro-osteogenic effects on osteoprogenitor cells via elaboration of extracellular vesicles (EV). These EV mediated effects were dependent on surface-associated tetraspanin expression, including CD9 and CD81. PSC derived EV stimulate bone defect repair, and do so via stimulation of skeletal progenitor cell proliferation, migration, and osteodifferentiation. In sum, PSC-EV represent an ‘off the shelf’ alternative for bone tissue engineering and regenerative medicine.
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Project description:Bone diseases profoundly affect patients, particularly the elderly, leading to severe health complications and disabilities. Osteoblasts play a crucial role in bone formation and are ideal candidates for treating bone diseases and engineering living materials. However, the stem and progenitor cells that give rise to osteoblasts, as well as osteoblasts themselves, exhibit dysfunction with aging. Although chemical reprogramming of fibroblasts into osteoblasts has been achieved, effective cell-based therapies or living materials have not been established in clinical practice. Here, we present a method using small molecules to achieve complete osteoblastic specification from human fibroblasts across all age groups. By targeting Wnt signaling pathways and modularizing small molecules and their combinations based on their effects on stage-specific genes, we optimized the temporal regulation of small molecules in reprogramming, achieving healthy induced osteoblasts(iOBs). The iOBs with traits of young native osteoblasts are ideal for forming transplantable tissue spheroids with improved survival, self-bone formation, and accelerated local angiogenesis in vivo, promoting effective bone defect repair. The material-free spheroids function as living, self-scaffolding building blocks for biofunctional constructs that reproduce tissue composition, maintain high cell density, and support matrix remodeling, offering a promising avenue for clinical autologous bone defect repair.
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Project description:Bone diseases profoundly affect patients, particularly the elderly, leading to severe health complications and disabilities. Osteoblasts play a crucial role in bone formation and are ideal candidates for treating bone diseases and engineering living materials. However, the stem and progenitor cells that give rise to osteoblasts, as well as osteoblasts themselves, exhibit dysfunction with aging. Although chemical reprogramming of fibroblasts into osteoblasts has been achieved, effective cell-based therapies or living materials have not been established in clinical practice. Here, we present a method using small molecules to achieve complete osteoblastic specification from human fibroblasts across all age groups. By targeting Wnt signaling pathways and modularizing small molecules and their combinations based on their effects on stage-specific genes, we optimized the temporal regulation of small molecules in reprogramming, achieving healthy induced osteoblasts(iOBs). The iOBs with traits of young native osteoblasts are ideal for forming transplantable tissue spheroids with improved survival, self-bone formation, and accelerated local angiogenesis in vivo, promoting effective bone defect repair. The material-free spheroids function as living, self-scaffolding building blocks for biofunctional constructs that reproduce tissue composition, maintain high cell density, and support matrix remodeling, offering a promising avenue for clinical autologous bone defect repair.
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