A Method for Lineage Tracing of Corneal Cells Using Multi-color Fluorescent Reporter Mice.
A Method for Lineage Tracing of Corneal Cells Using Multi-color Fluorescent Reporter Mice.
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DOI：
10.3791/53370
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发表时间：
2015-12-18
期刊：
影响因子：
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通讯作者：
Shalom-Feuerstein R
中科院分区：
文献类型：
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作者：
Amitai-Lange A;Berkowitz E;Altshuler A;Dbayat N;Nasser W;Suss-Toby E;Tiosano B;Shalom-Feuerstein R
谱系追踪实验定义了特定组织或生物体中细胞的起源、命运和行为。这项技术已经成功应用了几十年，揭示了发育生物学的开创性发现。最近，它被干细胞生物学家采用，以最小的实验干预来识别和跟踪不同的干细胞群。小鼠遗传学的最新发展，大量小鼠品系的可用性，以及荧光显微镜的进步允许使用商业上可用的工具，利用基本的专业知识，为各种组织直接设计强大的谱系追踪系统。我们最近利用这种强大的方法来探索干细胞的起源和命运在眼表使用R26 R-Confetti小鼠。该模型提供了一个多色遗传系统，用于以随机方式表达4个荧光基因。在这里，我们描述了这种方法的原则，并提供了一个适应性的协议设计谱系追踪实验，特别是角膜上皮以及其他组织。
Lineage tracing experiments define the origin, fate and behavior of cells in a specific tissue or organism. This technique has been successfully applied for many decades, revealing seminal findings in developmental biology. More recently, it was adopted by stem cell biologists to identify and track different stem cell populations with minimal experimental intervention. The recent developments in mouse genetics, the availability of a large number of mouse strains, and the advancements in fluorescent microscopy allow the straightforward design of powerful lineage tracing systems for various tissues with basic expertise, using commercially available tools. We have recently taken advantage of this powerful methodology to explore the origin and fate of stem cells at the ocular surface using R26R-Confetti mouse. This model offers a multi-color genetic system, for the expression of 4 fluorescent genes in a random manner. Here we describe the principles of this methodology and provide an adaptable protocol for designing lineage tracing experiments; specifically for the corneal epithelium as well as for other tissues.