Maintaining multipotent trunk neural crest stem cells as self-renewing crestospheres
Maintaining multipotent trunk neural crest stem cells as self-renewing crestospheres
复制标题
DOI：
10.1016/j.ydbio.2019.01.010
复制
发表时间：
2019-03-15
影响因子：
2.7
通讯作者：
Kerosuo, Laura
中科院分区：
文献类型：
--
作者：
Mohlin, Sofie;Kunttas, Ezgi;Kerosuo, Laura
神经脊细胞具有广泛的迁移和分化能力，这种能力随其轴向起源水平的不同而不同。然而，它们的暂时性限制了人们对其干细胞和自我更新特性的了解。虽然体外培养方法使颅神经脊细胞有可能保持为自我更新的多潜能脊球(Kerosuo等人，2015)，但这些相同的条件未能在干细胞状态下保存干神经脊细胞。在这里，我们优化了维持鸟类躯干冠球的培养条件，包括神经脊干细胞和祖细胞。我们的躯干衍生的眼球是多能的，并在几周内显示出自我更新的能力。与腹外侧神经管或中胚层命运的特征相比，躯干顶球的神经脊细胞标记物的表达增加。此外，与颅顶球相比，躯干冠球表达了更多的躯干神经脊丰富的标志物。最后，我们使用慢病毒转导作为一种工具来操纵躯干龟裂球中的基因表达。综上所述，这种方法能够在体外长期维持和操纵处于迁移前干细胞或早期祖细胞状态的多潜能干神经脊细胞。躯干棘球是探索神经脊骨粗细和血统决定以及伴随疾病的机制的宝贵资源。
Neural crest cells have broad migratory and differentiative ability that differs according to their axial level of origin. However, their transient nature has limited understanding of their stem cell and self-renewal properties. While an in vitro culture method has made it possible to maintain cranial neural crest cells as self-renewing multipotent crestospheres (Kerosuo et al., 2015), these same conditions failed to preserve trunk neural crest in a stem-like state. Here we optimize culture conditions for maintenance of avian trunk crestospheres, comprised of both neural crest stem and progenitor cells. Our trunk-derived crestospheres are multipotent and display self-renewal capacity over several weeks. Trunk crestospheres display elevated expression of neural crest cell markers as compared to those characteristic of ventrolateral neural tube or mesodermal fates. Moreover, trunk crestospheres express increased levels of trunk neural crest-enriched markers as compared to cranial crestospheres. Finally, we use lentiviral transduction as a tool to manipulate gene expression in trunk crestospheres. Taken together, this method enables long-term in vitro maintenance and manipulation of multipotent trunk neural crest cells in a premigratory stem or early progenitor state. Trunk crestospheres are a valuable resource for probing mechanisms underlying neural crest sternness and lineage decisions as well as accompanying diseases.