通过直接重编程小鼠成纤维细胞诱导成骨细胞。

Induction of Osteoblasts by Direct Reprogramming of Mouse Fibroblasts.

机构信息

Division of Endocrinology, Stanford University School of Medicine, Stanford, CA, USA.

出版信息

Methods Mol Biol. 2020;2155:201-212. doi: 10.1007/978-1-0716-0655-1_17.

Abstract

In the tissue culture dish, osteoblast cells can be derived from mesenchymal stem cells (MSCs) and pluripotent stem cells (PSCs) including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). However, differentiation of osteoblasts from PSCs is time-consuming and low yield. In contrast, we identified four osteogenic transcription factors, Runx2, Osx, Dlx5, and ATF4, that rapidly and efficiently reprogram mouse fibroblasts derived from 2.3 kb type I collagen promoter-driven green fluorescent protein (Col2.3GFP) transgenic mice into induced osteoblast cells (iOBs). iOBs exhibit osteoblast morphology, form mineralized nodules, and express Col2.3GFP and gene markers of osteoblast differentiation. Our method provides a robust system to rapidly generate appropriate and abundant osteoblast cells for osteogenesis and bone regeneration study.

摘要

在组织培养皿中，可以从间充质干细胞（MSCs）和多能干细胞（PSCs）中获得成骨细胞，包括胚胎干细胞（ESCs）和诱导多能干细胞（iPSCs）。然而，从 PSCs 中分化出成骨细胞是一个耗时且产量低的过程。相比之下，我们鉴定出了四个成骨转录因子，Runx2、Osx、Dlx5 和 ATF4，它们能够快速有效地将源自携带 2.3kb 类型 I 胶原启动子驱动的绿色荧光蛋白（Col2.3GFP）转基因小鼠的成纤维细胞重编程为诱导性成骨细胞（iOBs）。iOBs 表现出成骨细胞形态，形成矿化结节，并表达 Col2.3GFP 和成骨细胞分化的基因标志物。我们的方法提供了一个强大的系统，可以快速生成适当且丰富的成骨细胞，用于成骨和骨再生研究。