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.

遗传性骨骼疾病约占已记录的孟德尔疾病的 10%，并且与高经济负担有关。他们的研究需要成骨细胞，它们在调节骨组织的发育和维持中起关键作用。然而，骨组织并不总是可以从患者那里获得。我们开发了一种高效的基于血小板裂解物的方法，可将皮肤衍生的人成纤维细胞直接转分化为成骨细胞样细胞。我们通过在 mRNA 和蛋白质水平上检查转分化过程中成骨细胞特异性标志物的表达来广泛地表征我们的体外模型。转分化的成骨细胞样细胞显示一组成骨标志物的表达显着增加。还显示了矿物质沉积和 ALP 活性，证实了它们的成骨特性。RNA-seq 分析允许对转分化细胞转录组的变化进行全球研究。就显着上调的基因而言，转分化细胞与原代成纤维细胞分开聚集，表明不同的转录组谱；转分化成骨细胞也显示出与骨骼发育和骨矿化相关的基因表达显着富集。我们提出的体外模型可能有助于研究患者衍生细胞中成骨细胞依赖性疾病的前景。转分化成骨细胞也显示出与骨骼发育和骨矿化相关的基因表达显着富集。我们提出的体外模型可能有助于研究患者衍生细胞中成骨细胞依赖性疾病的前景。转分化成骨细胞也显示出与骨骼发育和骨矿化相关的基因表达显着富集。我们提出的体外模型可能有助于研究患者衍生细胞中成骨细胞依赖性疾病的前景。