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dc.contributor.author Walheim, Christopher C. en
dc.contributor.author Zanin, Juan Pablo en
dc.contributor.author De Bellard, Maria Elena en
dc.date.accessioned 2018-04-13T21:37:17Z
dc.date.available 2018-04-13T21:37:17Z
dc.date.issued 2012 en
dc.identifier.citation Walheim, C.C., Zanin, J.P., & de Bellard, M.E. (2012). Analysis of trunk neural crest cell migration using a modified Zigmond chamber assay. Journal of Visualized Experiments, 59, e3330. https://doi.org/10.3791/3330 en
dc.identifier.other 1940-087X en
dc.identifier.uri http://hdl.handle.net/10211.3/201966 en
dc.description.abstract Neural crest cells (NCCs) are a transient population of cells present in vertebrate development that emigrate from the dorsal neural tube (NT) after undergoing an epithelial-mesenchymal transition 1,2. Following EMT, NCCs migrate large distances along stereotypic pathways until they reach their targets. NCCs differentiate into a vast array of cell types including neurons, glia, melanocytes, and chromaffin cells 1-3. The ability of NCCs to reach and recognize their proper target locations is foundational for the appropriate formation of all structures containing trunk NCC-derived components 3. Elucidating the mechanisms of guidance for trunk NCC migration has therefore been a matter of great significance. Numerous molecules have been demonstrated to guide NCC migration 4. For instance, trunk NCCs are known to be repelled by negative guidance cues such as Semaphorin, Ephrin, and Slit ligands 5-8. However, not until recently have any chemoattractants of trunk NCCs been identified 9. Conventional in vitro approaches to studying the chemotactic behavior of adherent cells work best with immortalized, homogenously distributed cells, but are more challenging to apply to certain primary stem cell cultures that initially lack a homogenous distribution and rapidly differentiate (such as NCCs). One approach to homogenize the distribution of trunk NCCs for chemotaxis studies is to isolate trunk NCCs from primary NT explant cultures, then lift and replate them to be almost 100% confluent. However, this plating approach requires substantial amounts of time and effort to explant enough cells, is harsh, and distributes trunk NCCs in a dissimilar manner to that found in in vivo conditions. Here, we report an in vitro approach that is able to evaluate chemotaxis and other migratory responses of trunk NCCs without requiring a homogenous cell distribution. This technique utilizes time-lapse imaging of primary, unperturbed trunk NCCs inside a modified Zigmond chamber (a standard Zigmond chamber is described elsewhere10). By exposing trunk NCCs at the periphery of the culture to a chemotactant gradient that is perpendicular to their predicted natural directionality, alterations in migratory polarity induced by the applied chemotactant gradient can be detected. This technique is inexpensive, requires the culturing of only two NT explants per replicate treatment, avoids harsh cell lifting (such as trypsinization), leaves trunk NCCs in a more similar distribution to in vivo conditions, cuts down the amount of time between explantation and experimentation (which likely reduces the risk of differentiation), and allows time-lapse evaluation of numerous migratory characteristics. en
dc.language.iso en en
dc.publisher Journal of Visualized Experiments en
dc.relation.uri https://doi.org/10.3791/3330 en
dc.subject neural crest cells en
dc.subject vertebrate en
dc.subject cell migration en
dc.title Analysis of trunk neural crest cell migration using a modified Zigmond chamber assay en
dc.type Presentation en
dc.identifier.orcid orcid.org/0000-0001-9881-0450 en


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  • De Bellard, Maria [27]
    Collection of scholarship submitted by Dr. Maria De Bellard, Department of Biology
  • Faculty Publications [2384]
    Collection of scholarship submitted by CSUN faculty

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