Recapitulating early development of mouse musculoskeletal precursors of the paraxial mesoderm in vitro
Recapitulating early development of mouse musculoskeletal precursors of the paraxial mesoderm in vitro
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DOI：
10.1242/dev.157339
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发表时间：
2018-03-01
期刊：
影响因子：
4.6
通讯作者：
Pourquie, Olivier
中科院分区：
文献类型：
--
作者：
Chal, Jerome;Al Tanoury, Ziad;Pourquie, Olivier
身体骨骼肌源自近轴中胚层，形成于胚胎的后部区域。使用微阵列，我们表征了新型小鼠前体中胚层（PSM）标记，并表明，与 PSM 的突然转录组重组不同，神经管分化伴随着渐进的转录组变化。使用小鼠和人类多能干细胞可以在体外有效地重现早期近轴中胚层分化阶段。虽然单独激活 Wnt 可以诱导后部 PSM 标记，但获得特定的 PSM 命运并有效分化为前部 PSM Pax3(+) 身份进一步需要 BMP 抑制，以防止祖细胞漂移至侧板中胚层命运。当移植到受伤的成年肌肉中时，这些前体细胞会产生大量未成熟的肌纤维。此外，将这些小鼠 PSM 样细胞暴露于短暂的 FGF 抑制步骤，然后在含马血清的培养基中培养，可以有效地重现生肌程序，以生成肌管和相关的 Pax7(+) 细胞。与无血清方案相比，该方案可改善小鼠肌纤维的体外分化和成熟，并使肌源细胞融合和卫星细胞分化的研究成为可能。
Body skeletal muscles derive from the paraxial mesoderm, which forms in the posterior region of the embryo. Using microarrays, we characterize novel mouse presomitic mesoderm (PSM) markers and show that, unlike the abrupt transcriptome reorganization of the PSM, neural tube differentiation is accompanied by progressive transcriptome changes. The early paraxial mesoderm differentiation stages can be efficiently recapitulated in vitro using mouse and human pluripotent stem cells. While Wnt activation alone can induce posterior PSM markers, acquisition of a committed PSM fate and efficient differentiation into anterior PSM Pax3(+) identity further requires BMP inhibition to prevent progenitors from drifting to a lateral plate mesoderm fate. When transplanted into injured adult muscle, these precursors generated large numbers of immature muscle fibers. Furthermore, exposing these mouse PSM-like cells to a brief FGF inhibition step followed by culture in horse serum-containing medium allows efficient recapitulation of the myogenic program to generate myotubes and associated Pax7(+) cells. This protocol results in improved in vitro differentiation and maturation of mouse muscle fibers over serum-free protocols and enables the study of myogenic cell fusion and satellite cell differentiation.