| Isolation of Mouse Cardiac Neural Crest Cells and Their Differentiation into Smooth Muscle Cells
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Author:
Date:
2017-09-05
[Abstract] Cardiac neural crest cells (CNCCs) originate at the dorsal edge of the neural tube between the otic pit and the caudal edge of the 3rd somite, and migrate into the pharyngeal arches and the heart. We have shown that fibronectin (Fn1) plays an important role in the development of the CNCC by regulating the differentiation of CNCCs into vascular smooth muscle cells around pharyngeal arch arteries (Wang and Astrof, 2016). This protocol describes the isolation of CNCCs from the neural tube and from the caudal pharyngeal arches, and the differentiation of neural crest-derived cells into smooth muscle cells. This protocol was adapted from (Newgreen and Murphy, 2000; Pfaltzgraff et al., 2012).
[摘要] 心脏神经嵴细胞（CNCC）起源于神经管的背部边缘，位于第3个体节的耳穴和尾缘之间，并迁移到咽弓和心脏。 我们已经表明，纤连蛋白（Fn1）通过调节CNCCs到咽弓动脉周围的血管平滑肌细胞的分化，在CNCC的发展中起重要作用（Wang and Astrof，2016）。 该方案描述了CNCC与神经管和尾尾弓的分离，以及神经嵴衍生细胞分化成平滑肌细胞。 该方案从（Newgreen和Murphy，2000; Pfaltzgraff等人，2012）改编。
【背景】以前发表的方案描述了从神经管分离神经嵴细胞。然而，在耳孔和第三体细胞之间的神经管区域中的神经嵴细胞包括有助于许多不同细胞类型的神经嵴细胞群体;例如，迷走神经嵴细胞也来自该区域。在该方案中，我们修改了用于分离心脏神经嵴细胞的常规方法。而不是使用神经管，我们在胚胎期（E）9.5（22-25个体节期）使用尾部咽部弓形区。这是在将心脏神经嵴细胞分化为血管平滑肌细胞之前。神经嵴培养物通常含有污染性间充质细胞，通常表达平滑肌基因。为了鉴定神经嵴衍生细胞，我们从以下交叉产生的胚胎中分离出神经嵴细胞：Fn1flox / flox; ROSAmTmG / mTmG雌性小鼠×Fn1 +/-;Tfap2αIRESCre/ ...
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| Clonal Culture of Mouse Liver Progenitor Cells
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Author:
Date:
2015-10-20
[Abstract] Liver stem/progenitor cells (LPCs) are defined as bipotential cells differentiating into both hepatocytes and cholangiocytes. For analyzing their differentiation potential, clonal culture has been used for LPCs isolated by a cell sorter. In addition, we can use the culture to assess functions of target genes on differentiation potential of LPCs. This protocol describes the process of cell isolation and colony assay to examine proliferative and differentiation potential of LPCs.
[摘要] 肝干/祖细胞（LPC）被定义为分化成肝细胞和胆管细胞的双能细胞。 为了分析其分化潜力，克隆培养已经用于通过细胞分选仪分离的LPC。 此外，我们可以使用文化来评估目标基因的功能分化潜力的LPCs。 该协议描述细胞分离和集落分析的过程，以检查增殖和分化潜力的LPCs。
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