Osteogenic transdifferentiation of primary human fibroblasts to osteoblast-like cells with human platelet lysate

Abstract

Inherited bone disorders account for about 10% of documented Mendelian disorders and are associated with high financial burden. Their study requires osteoblasts which play a critical role in regulating the development and maintenance of bone tissue. However, bone tissue is not always available from patients. We developed a highly efficient platelet lysate-based approach to directly transdifferentiate skin-derived human fibroblasts to osteoblast-like cells. We extensively characterized our in vitro model by examining the expression of osteoblast-specific markers during the transdifferentiation process both at the mRNA and protein level. The transdifferentiated osteoblast-like cells showed significantly increased expression of a panel of osteogenic markers. Mineral deposition and ALP activity were also shown, confirming their osteogenic properties. RNA-seq analysis allowed the global study of changes in the transcriptome of the transdifferentiated cells. The transdifferentiated cells clustered separately from the primary fibroblasts with regard to the significantly upregulated genes indicating a distinct transcriptome profile; transdifferentiated osteoblasts also showed significant enrichment in gene expression related to skeletal development and bone mineralization. Our presented in vitro model may potentially contribute to the prospect of studying osteoblast-dependent disorders in patient-derived cells.

Vitamin D Attenuates Fibrotic Properties of Fibrous Dysplasia-Derived Cells for the Transit towards Osteocytic Phenotype

“…Additionally, we demonstrated that 1,25(OH)2D3 treatment enhances the mineralizing capacities of FD-derived cells, as evidenced by increased ALP activity and ARS staining intensity. These findings suggest that 1,25(OH)2D3 promotes osteogenic differentiation and mineralization in FD-derived While traditionally fibroblasts were not considered direct precursors of osteoblasts, and vice versa, emerging evidence suggests that they possess differentiation plasticity, allowing for transdifferentiation processes [31]. This phenomenon is evident in certain pathological contexts like ossification of the posterior longitudinal ligament (OPLL), where fibroblasts within ligament tissue differentiate into osteoblast-like cells, contributing to ectopic bone formation [32,33].…”

Analysis of intercellular communication in the osteosarcoma microenvironment based on single cell sequencing data

“…It's well known that multiple important cell types in mammalian bone tissue have a mesenchymal origin, including fibroblasts, osteoblasts, chondroblasts. These cells can undergo transdifferentiation under specific conditions or environments, such as transdifferentiation of chondrocytes into osteoblasts [19] , [35] , transdifferentiation of fibroblasts into osteoblasts [36] , [37] , [38] . I-Ning E Wang et al [39] found that the interaction of osteoblasts and fibroblasts lead to cell trans -differentiation and fibrocartilage formation in a vitro co-culture system.…”

A Systematic Review of the Evidence of Hematopoietic Stem Cell Differentiation to Fibroblasts

“…The ability of hematopoietic stem cells to differentiate into (myo)fibroblasts and other cell types may have significant implications for a range of pathophysiological processes. Examples include organ fibrosis, dystrophic epidermolysis bullosa, leptin deficiency and even rare bone diseases such as fibrodysplasia ossificans progressiva in which the process of heterotopic ossification is possibly facilitated through fibroblasts [ 1 , 113 , 114 , 115 , 116 , 117 ]. It is plausible that the properties of HSCs may potentially affect disease presentation.…”