Isolation, Purification, and Culture of Embryonic Melanoblasts from Green Fluorescent Protein-expressing Reporter Mice.

Melissa Crawford, Kevin Barr, Lina Dagnino
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Abstract

In this article, we provide a method to isolate embryonic melanoblasts from reporter mouse strains. The mice from which these cells are isolated are bred into the ROSA26mT/mG reporter background, which results in green fluorescent protein (GFP) expression in the targeted melanoblast population. These cells are isolated and purified by fluorescence-activated cell sorting using GFP fluorescence. We also provide a method to culture the purified melanoblasts for further analysis. This method yields > 99% purity melanoblasts specifically targeted, and can be used for a variety of studies, including gene expression, clonogenic experiments, and biological assays, such as viability, capacity for directional migration, or differentiation into melanin-producing melanocytic cells.

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表达绿色荧光蛋白的报告小鼠胚胎黑色素母细胞的分离、纯化和培养。
在本文中,我们提供了一种从报告鼠株中分离胚胎黑素母细胞的方法。将分离出这些细胞的小鼠培育到ROSA26mT/mG报告细胞背景中,结果在目标黑色素细胞群体中表达绿色荧光蛋白(GFP)。这些细胞是用荧光激活的细胞分选技术分离纯化的。我们还提供了一种培养纯化成黑素细胞的方法,以便进一步分析。该方法可获得纯度> 99%的特异性黑素母细胞,并可用于多种研究,包括基因表达、克隆实验和生物分析,如活力、定向迁移能力或分化为产生黑色素的黑素细胞。
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