Fluorescence-activated cell sorting of EGFP-labeled neural crest cells from murine embryonic craniofacial tissue.
Fluorescence-activated cell sorting of EGFP-labeled neural crest cells from murine embryonic craniofacial tissue.
复制标题
EGFP标记的神经rest细胞的荧光激活细胞从鼠类胚胎颅面组织中。
DOI：
10.1155/jbb.2005.232
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发表时间：
2005
影响因子：
--
通讯作者：
Pisano, MM
中科院分区：
文献类型：
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作者：
Singh, S;Bhattacherjee, V;Mukhopadhyay, P;Worth, CA;Wellhausen, SR;Warner, CP;Greene, RM;Pisano, MM
在胚胎发生的早期阶段，已知多能神经嵴细胞（NCC）从神经褶皱迁移到胚胎中的多个目标部位，在那里它们分化成各种衍生物，包括软骨、骨、结缔组织、黑素细胞、胶质细胞和周围神经系统的神经元。获得纯NCC群体的能力对于神经嵴诱导、迁移和/或分化的分子分析至关重要。将Wnt1-Cre和Z/EG转基因小鼠系杂交得到的后代中，Wnt1-Cre转基因仅在NCC中激活了EGFP的永久表达。本报告展示了一种流式细胞术方法来分类和分离egfp标记的NCC群体。通过TaqMan定量实时聚合酶链式反应（QRT-PCR）检测已知标记基因的表达，确认所选神经嵴细胞的身份。本报告中描述的分子策略提供了一种从纯NCC群体中提取完整RNA的方法，从而可以分析对发育至关重要的特定胚胎前体细胞群体中的基因表达。
During the early stages of embryogenesis, pluripotent neural crest cells (NCC) are known to migrate from the neural folds to populate multiple target sites in the embryo where they differentiate into various derivatives, including cartilage, bone, connective tissue, melanocytes, glia, and neurons of the peripheral nervous system. The ability to obtain pure NCC populations is essential to enable molecular analyses of neural crest induction, migration, and/or differentiation. Crossing Wnt1-Cre and Z/EG transgenic mouse lines resulted in offspring in which the Wnt1-Cre transgene activated permanent EGFP expression only in NCC. The present report demonstrates a flow cytometric method to sort and isolate populations of EGFP-labeled NCC. The identity of the sorted neural crest cells was confirmed by assaying expression of known marker genes by TaqMan Quantitative Real-Time Polymerase Chain Reaction (QRT-PCR). The molecular strategy described in this report provides a means to extract intact RNA from a pure population of NCC thus enabling analysis of gene expression in a defined population of embryonic precursor cells critical to development.