Published: Vol 3, Iss 15, Aug 5, 2013 DOI: 10.21769/BioProtoc.840 Views: 11054
Reviewed by: Fanglian He
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Abstract
Extravillous trophoblast (EVT) migration and invasion through the decidualized endometrium is essential to successful placentation. SGHPL-4 cells, an EVT cell line derived from first trimester placenta, is a widely used model of cytotrophoblast differentiation into an invasive phenotype. Here we describe a quantitative cell migration assay that can be modified to also measure cell invasion. SGHPL-4 cells were seeded into BD Fluoroblok cell culture inserts constructed with an 8 μm porous membrane and allowed to migrate towards epidermal growth factor, a known chemoattractant for EVTs. To assess EVT invasion, Fluoroblok inserts were first coated with Matrigel, a basement membrane matrix. SGHPL-4 cells were labeled with calcein AM and cells that had invaded and/or migrated across the membrane were quantified by a bottom-reading fluorescence plate reader. The advantage of the Fluoroblok inserts over other migration/invasion assays is that they allow nondestructive detection of migrated cells.
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Acknowledgments
This protocol is adapted from Angelova et al. (2012).
References
Article Information
Copyright
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Angelova, M., Machado, H. L., Swan, K. F., Morris, C. and Sullivan, D. E. (2013). Extravillous Trophoblast Migration and Invasion Assay. Bio-protocol 3(15): e840. DOI: 10.21769/BioProtoc.840.
Category
Cell Biology > Cell movement > Cell migration
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