Abstract
BackgroundAlterations in cell migration are a hallmark of cancer cell invasion and metastasis. In vitro assays commonly used to study cell migration, including the scratch wound healing assay, Boyden chamber assay, and newly developed advanced systems with microfluidics, each have several disadvantages.FindingsHere we describe an easy and cost-effective in vitro assay for cell migration employing cloning rings to create gaps in the cell monolayer (“ring cell migration assay”). The assay was used to quantitate innate differences in cell motility and the effect of various extracellular matrix proteins on migration of five cancer cell lines: U87 and U251N glioma cells, MDA-MB-231and MCF-7 breast cancer cells, and HeLa cervical cancer cells. Interestingly, collagen was a general promoter of cell migration for all five cancer cell lines, without affecting cell proliferation.ConclusionsTaken together, the ring cell migration assay is an easy, convenient and cost-effective assay to study cell migration in vitro.
Highlights
Alterations in cell migration are a hallmark of cancer cell invasion and metastasis
Materials and methods Cell culture and reagents Two glioma cell lines U87 and U251N, two breast cancer cell lines MDA-MB-231 and MCF-7, and HeLa cervical cancer cells were obtained from American Type Cell Collection (ATCC) (Manassas, VA) and routinely maintained in Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS) supplemented with 100 μg/ml of penicillin-streptomycin
We found that cell confluency did not significantly affect the migration rate of cells and that the difference in cell motility of the different cell lines was independent of cell confluency; for example, U87 cells at lower confluency migrated faster than U251N cells did at higher confluency (Figure 2), suggesting that cell motility is an intrinsic property of migrating cells
Summary
Metastasis is the leading cause of mortality for patients suffering from cancer. During metastasis, abnormal cells with genomic heterogeneity may undergo epithelial-tomesenchymal transition (EMT) and phenotypically exhibit increased motility and invasiveness as compared to normal cells [1,2,3]. ECM plays critical roles in cancer cell invasion and metastasis [7,8,9,10] It is mostly unknown how various ECM proteins affect cell migration of different tissuespecific cancer cell types. There are several conventional in vitro assays for studying cancer cell migration, including scratch wound healing assay [13] and Boyden chamber assay [14]. While these assays have advantages either in ease of performance (scratch assay) or in mimicking in vivo chemoattractant gradients for cell migration (Boyden assay), they have many disadvantages. We tested five cancer cell lines of different tissue origin to verify the assay’s ability to distinguish differences in cancer cell motility and to analyze the effect of various extracellular matrix proteins on cancer cell migration
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
