Statistical analysis of cell migration in 3D using the anisotropic persistent random walk model

Wu, Pei Hsun; Giri, Anjil; Wirtz, Denis

doi:10.1038/nprot.2015.030

Cited by 99 publications

(140 citation statements)

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“…1 A–N ). Trajectories of cells ( x , y coordinates) were transformed into MSDs and evaluated (25, 36). On 2D substrates, the MSDs evaluated at 1 h were reduced by 41 (0.1 mg/ml) and 52% (1 mg/ml) for nocodazole-treated cells and by 8 (1 pM), 84 (0.1 nM), and 91% (100 nM) for taxol-treated cells compared with control cells (Fig.…”

Section: Resultsmentioning

confidence: 99%

EB1 and cytoplasmic dynein mediate protrusion dynamics for efficient 3‐dimensional cell migration

et al. 2018

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Microtubules have long been implicated to play an integral role in metastatic disease, for which a critical step is the local invasion of tumor cells into the 3-dimensional (3D) collagen-rich stromal matrix. Here we show that cell migration of human cancer cells uses the dynamic formation of highly branched protrusions that are composed of a microtubule core surrounded by cortical actin, a cytoskeletal organization that is absent in cells on 2-dimensional (2D) substrates. Microtubule plus-end tracking protein End-binding 1 and motor protein dynein subunits light intermediate chain 2 and heavy chain 1, which do not regulate 2D migration, critically modulate 3D migration by affecting RhoA and thus regulate protrusion branching through differential assembly dynamics of microtubules. An important consequence of this observation is that the commonly used cancer drug paclitaxel is 100-fold more effective at blocking migration in a 3D matrix than on a 2D matrix. This work reveals the central role that microtubule dynamics plays in powering cell migration in a more pathologically relevant setting and suggests further testing of therapeutics targeting microtubules to mitigate migration.—Jayatilaka, H., Giri, A., Karl, M., Aifuwa, I., Trenton, N. J., Phillip, J. M., Khatau, S., Wirtz, D. EB1 and cytoplasmic dynein mediate protrusion dynamics for efficient 3-dimensional cell migration.

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Section: Resultsmentioning

confidence: 99%

EB1 and cytoplasmic dynein mediate protrusion dynamics for efficient 3‐dimensional cell migration

et al. 2018

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“…Recently, Wu et al developed a variation on the persistent random walk model, taking into account that certain cell types have a preferred direction of migration. 46, 47 The anisotropic random walk model (APRW) is well suited for our experiments, as migration on microprinted lines and in microchannels represents the extreme case of this model by exclusively allowing migration along one axis. Two descriptors of the APRW model, namely, primary persistence time and total diffusivity, were calculated here.…”

Section: Resultsmentioning

confidence: 99%

Time-lapse lens-free imaging of cell migration in diverse physical microenvironments

et al. 2016

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Time-lapse imaging of biological samples is important for understanding complex (patho)physiological processes. A growing number of point-of-care biomedical assays rely on real-time imaging of flowing or migrating cells. However, the cost and complexity of integrating experimental models simulating physiologically relevant microenvironments with bulky imaging systems that offer sufficient spatiotemporal resolution limit the use of time-lapse assays in research and clinical settings. This paper introduces a compact and affordable lens-free imaging (LFI) device based on the principle of coherent in-line, digital holography for time-lapse cell migration assays. The LFI device combines single-cell resolution (1.2 μm) with a large field of view (6.4 × 4.6 mm2), thus rendering it ideal for high-throughput applications and removing the need for expensive and bulky programmable motorized stages. The set-up is so compact that it can be housed in a standard cell culture incubator, thereby avoiding custom-built stage top incubators. LFI is thoroughly benchmarked against conventional live-cell phase contrast microscopy for random cell motility on two-dimensional (2D) surfaces and confined migration on 1D-microprinted lines and in microchannels using breast adenocarcinoma cells. The quality of the results obtained by the two imaging systems is comparable, and they reveal that cells migrate more efficiently upon increasing confinement. Interestingly, assays of confined migration more readily distinguish the migratory potential of metastatic MDA-MB-231 cells from non-metastatic MCF7 cells relative to traditional 2D migration assays. Altogether, this single-cell migration study establishes LFI as an elegant and useful tool for live-cell imaging.

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“…The images were analyzed using Imaris spot analysis software (Imaris 8.1; Bitplane) to track the time-dependent mobility. The 3D migration analysis was performed following the strategy developed by Wirtz and coworkers (29,30). A minimum of 100 individual cells at each point were tracked to generate x, y, and z coordinates at each time point.…”

Section: Methodsmentioning

confidence: 99%

“…Graphed data are presented as average ± SD. Significance levels were set at *P < 0.05,^P < 0.01, and # P < 0.001 (29,30).…”

Section: Methodsmentioning

confidence: 99%

“…By choosing the time frame with the most visible cells we could maximize the sample size of cells that could be analyzed. Finally, velocity and speed profiles, MSDs, and trajectory plots were calculated using code adapted from Wirtz and coworkers (29,30) for triplicate tracking trials (n = 3). The statistical analysis was performed using MATLAB (Mathworks, Inc.) to calculate the mean, SD, and SE of the mean.…”

Section: Methodsmentioning

confidence: 99%

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Intratumoral oxygen gradients mediate sarcoma cell invasion

Lewis

Park

Tang

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

104

117

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Hypoxia is a critical factor in the progression and metastasis of many cancers, including soft tissue sarcomas. Frequently, oxygen (O 2 ) gradients develop in tumors as they grow beyond their vascular supply, leading to heterogeneous areas of O 2 depletion. Here, we report the impact of hypoxic O 2 gradients on sarcoma cell invasion and migration. O 2 gradient measurements showed that large sarcoma mouse tumors (>300 mm 3 ) contain a severely hypoxic core [≤0.1% partial pressure of O 2 (pO 2 )] whereas smaller tumors possessed hypoxic gradients throughout the tumor mass (0.1-6% pO 2 ). To analyze tumor invasion, we used O 2 -controllable hydrogels to recreate the physiopathological O 2 levels in vitro. Small tumor grafts encapsulated in the hydrogels revealed increased invasion that was both faster and extended over a longer distance in the hypoxic hydrogels compared with nonhypoxic hydrogels. To model the effect of the O 2 gradient accurately, we examined individual sarcoma cells embedded in the O 2 -controllable hydrogel. We observed that hypoxic gradients guide sarcoma cell motility and matrix remodeling through hypoxia-inducible factor-1α (HIF-1α) activation. We further found that in the hypoxic gradient, individual cells migrate more quickly, across longer distances, and in the direction of increasing O 2 tension. Treatment with minoxidil, an inhibitor of hypoxia-induced sarcoma metastasis, abrogated cell migration and matrix remodeling in the hypoxic gradient. Overall, we show that O 2 acts as a 3D physicotactic agent during sarcoma tumor invasion and propose the O 2 -controllable hydrogels as a predictive system to study early stages of the metastatic process and therapeutic targets.hydrogel | sarcoma | hypoxia | gradients | migration S oft tissue sarcomas are a heterogeneous group of malignant cancers derived from transformed cells of mesenchymal origin (1, 2). Approximately 13,000 new cases per year are diagnosed in the United States alone, with 25-50% of patients developing recurrent and metastatic disease (3-5). Current clinical data suggest that undifferentiated pleomorphic sarcoma (UPS) is one of the most aggressive sarcoma subtypes, which frequently results in lethal pulmonary metastases that are insensitive to radio/chemotherapy. It has recently become apparent that sarcoma progression and metastasis are regulated by microenvironmental cues, such as extracellular matrix (ECM) remodeling, stiffness modulation, cellto-cell/matrix interactions, signaling factors, and spatial gradients (6-8). Of all these factors, low intratumoral oxygen (O 2 ; hypoxia) is most dramatically associated with pulmonary metastasis and poor clinical outcomes (9, 10).Intratumoral hypoxia occurs when the partial pressure of O 2 (pO 2 ) falls below 5%, and it is a commonly observed feature of many sarcomas. Regional hypoxia develops as rapidly growing tumors outstrip their blood supply and as a consequence of aberrant tumor angiogenesis. As a result, O 2 gradients develop throughout the growing tumor. Tumor hypoxia promotes c...

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Statistical analysis of cell migration in 3D using the anisotropic persistent random walk model

Cited by 99 publications

References 35 publications

EB1 and cytoplasmic dynein mediate protrusion dynamics for efficient 3‐dimensional cell migration

EB1 and cytoplasmic dynein mediate protrusion dynamics for efficient 3‐dimensional cell migration

Time-lapse lens-free imaging of cell migration in diverse physical microenvironments

Intratumoral oxygen gradients mediate sarcoma cell invasion

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