Measuring Process Dynamics and Nuclear Migration for Clones of Neural Progenitor Cells
Neural stem and progenitor cells (NPCs) generate processes that extend from the cell body in a dynamic manner. The NPC nucleus migrates along these processes with patterns believed to be tightly coupled to mechanisms of cell cycle regulation and cell fate
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Department of Electrical and Computer Engineering, Drexel University, Philadelphia, PA, USA [email protected] 2 Neural Stem Cell Institute, Rensselaer, NY, USA
Abstract. Neural stem and progenitor cells (NPCs) generate processes that extend from the cell body in a dynamic manner. The NPC nucleus migrates along these processes with patterns believed to be tightly coupled to mechanisms of cell cycle regulation and cell fate determination. Here, we describe a new segmentation and tracking approach that allows NPC processes and nuclei to be reliably tracked across multiple rounds of cell division in phase-contrast microscopy images. Results are presented for mouse adult and embryonic NPCs from hundreds of clones, or lineage trees, containing tens of thousands of cells and millions of segmentations. New visualization approaches allow the NPC nuclear and process features to be effectively visualized for an entire clone. Significant differences in process and nuclear dynamics were found among type A and type C adult NPCs, and also between embryonic NPCs cultured from the anterior and posterior cerebral cortex. Keywords: Neural stem cells · Neural progenitor cells · Stem cell processes · Segmentation · Tracking · Lineaging · Stem cell process dynamics · Interkinetic nuclear migration
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Introduction
Neural progenitor cells (NPCs) play a key role in the generation and maintenance of the nervous system. NPCs are proliferative, undergoing mitosis to generate daughter cells that are genetic copies of the parent. As this process repeats, a family tree, or clone of related cells develops. NPCs are also migratory, with cells moving as needed to form and maintain the functionally, spatially and morphologically distinct components of the nervous system. Individual NPCs exhibit a complex cellular morphology, with rapidly forming and changing cellular processes or protrusions [1]. These NPC processes may be used to attach to different regional structures such as the apical or basal surface of the neuroepithelium during development [2], and may also be used to explore the environment. During the non-mitotic, or interkinetic portion of c Springer International Publishing Switzerland 2016 G. Hua and H. J´ egou (Eds.): ECCV 2016 Workshops, Part I, LNCS 9913, pp. 291–305, 2016. DOI: 10.1007/978-3-319-46604-0 21
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the cell cycle, NPC nuclei undergo distinct motion along the processes. This interkinetic nuclear migration (INM) has been observed in a wide variety of vertebrate cell types [3], and has been associated with the cell fate, or type of daughter cells that will be produced post-mitosis [4]. The relationship between INM and mechanisms controlling cell cycle and mitosis is an important open question [5,6]. Fluorescence microscopy is most commonly used to visualize INM. Because INM generally involves the spatial patterning or organization of a 3-D structure such as the cortex or retina, 3-D fluorescence microscopy, either confocal or multiphoton, is a way to observe not only the INM, but also the
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