Programmed subcellular release for studying the dynamics of cell detachment

Wildt, Bridget; Wirtz, Denis; Searson, Peter C.

doi:10.1038/nmeth.1299

Cited by 42 publications

(40 citation statements)

References 16 publications

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“…Recently, this has been achieved in predefined geometries on the surface of glass with an array of gold features that electrochemically releases RGD. 28 In addition, adhesive ligands have been photochemically exposed on glass via single and two-photon irradiation of photoactive SAMs. 25, 26, 37 Here, two-photon induced photodegradation of PEGdiPDA hydrogels offers a soft substrate with tunable elasticity and user-defined geometrical and temporal control of the ECM landscape at both the surface of 2D hydrogel scaffolds and within 3D hydrogels.…”

Section: Resultsmentioning

confidence: 99%

“…4b), where ΔA( t ) is the change in cell area at time t (A 0 − A( t )) and A 0 is the initial cell area, similar to the approach of Wildt et al 28 The following expression that describes the relationship between cell area and retraction time was fit to the data corresponding to each retracting cell:

\frac{Δ A (t)}{A_{0}} = \frac{Δ A_{max}}{A_{0}} [1 - exp (- \frac{t - t_{0}}{τ_{c}})]

where ΔA max is the maximum change in cell area; t 0 is the induction time prior to retraction during which A ≅ A 0 ; and τ c is the characteristic retraction time for the cell. 28 The fit to this model (Fig. 4b) suggests that the cell is under tension prior to detachment and undergoes damped retraction.…”

Section: Resultsmentioning

confidence: 99%

“…Using this approach we quantified subcellular detachment of MSCs from soft materials, which was observed to be ~6 fold slower than detachment from stiffer substrates. 28 PEGdiPDA hydrogels combined with two-photon induced erosion are useful tools for studying cell adhesion and detachment to and from soft substrates with tunable elasticity, and these materials should facilitate fundamental investigations of the dynamics of cell mechanobiology in both 2D and 3D culture.…”

Section: Discussionmentioning

confidence: 99%

“…24 In complementary approaches, investigators have developed responsive material substrates that can trigger changes in surface chemistry to temporally manipulate adhesive ligand presentation 25, 26 and cell attachment. 27, 28 For example, Wildt et al presented a strategy to induce subcellular detachment by electrochemically releasing the fibronectin-derived adhesive ligand RGD from micropatterned gold features on glass and reported quantitative analysis of the detachment dynamics. 28 This technique allows the user to control adhesion dynamically on stiff substrates in predefined geometries.…”

Section: Introductionmentioning

confidence: 99%

“…27, 28 For example, Wildt et al presented a strategy to induce subcellular detachment by electrochemically releasing the fibronectin-derived adhesive ligand RGD from micropatterned gold features on glass and reported quantitative analysis of the detachment dynamics. 28 This technique allows the user to control adhesion dynamically on stiff substrates in predefined geometries.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Controlled two-photon photodegradation of PEG hydrogels to study and manipulate subcellular interactions on soft materials

et al. 2010

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Cell adhesion and detachment to and from the extracellular matrix (ECM) are critical regulators of cell function and fate due to the exchange of mechanical signals between the cell and its microenvironment. To study this cell mechanobiology, researchers have developed several innovative methods to investigate cell adhesion in vitro; however, most of these culture platforms are unnaturally stiff or static. To better capture the soft, dynamic nature of the ECM, we present a PEG-based hydrogel in which the context and geometry of the extracellular space can be precisely controlled in situ via two-photon induced erosion. Here, we characterize the two-photon erosion process, demonstrate its efficacy in the presence of cells, and subsequently exploit it to induce subcellular detachment from soft hydrogels. A working space was established for a range of laser powers required to induce complete erosion of the gel, and these data are plotted with model predictions. From this working space, two-photon irradiation parameters were selected for complete erosion in the presence of cells. Micron-scale features were eroded on and within a gel to demonstrate the resolution of patterning with these irradiation conditions. Lastly, two-photon irradiation was used to erode the material at the cell-gel interface to remove cell adhesion sites selectively, and cell retraction was monitored to quantify the mesenchymal stem cell (MSC) response to subcellular detachment from soft materials.

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Section: Resultsmentioning

confidence: 99%

\frac{Δ A (t)}{A_{0}} = \frac{Δ A_{max}}{A_{0}} [1 - exp (- \frac{t - t_{0}}{τ_{c}})]

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Controlled two-photon photodegradation of PEG hydrogels to study and manipulate subcellular interactions on soft materials

et al. 2010

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Harvesting of Living Cell Sheets by the Dynamic Generation of Diffractive Photothermal Pattern on PEDOT

Heo

Han

et al. 2017

Adv Funct Materials

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Near infrared (NIR) photothermal pattern on conductive polymer film enables unique approaches to harvest large‐area cell sheets with various patterns without the use of patterned culture dish. The NIR photothermal pattern is generated from a patterned optical lens (POL), which creates a dynamic near IR light pattern and the corresponding photothermal pattern (PTP) on the polymer film. The POL is prepared from transparent polydimethylsiloxane designed to generate various light patterns. The PTPs allow a noninvasive harvest of cultured cells as an intact living cell sheet with a high harvesting efficiency (ηcell > 100). Various PTPs are generated by the diffraction of NIR light through POLs having different micropatterns, which afford cell sheets with a desired pattern without changing the original cell morphology at cultured state. Furthermore, a large‐area living cell sheet is obtained with a detached area larger than 19 cm2, which is the largest living cell sheet up to date. Further optical engineering of the harvesting system allows multiple productions of cell sheets with one dose of light. It is possible to harvest cell sheets not only from human fibroblast cells but also from human adipose‐derived stem cells, indicating that the method can be applied to engineer various cells.

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In Situ Subcellular Detachment of Cells Using a Cell‐Friendly Photoresist and Spatially Modulated Light

et al. 2019

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Dynamic adhesion and detachment of subcellular regions occur during cell migration, thus a technique allowing precise control of subcellular detachment of cells will be useful for cell migration study. Previous methods for cell detachment were developed either for harvesting cells or cell sheets attached on surfaces with low resolution patterning capability, or for detaching subcellular regions located on predefined electrodes. In this paper, a method that allows in situ subcellular detachment of cells with ≈1.5 µm critical feature size while observing cells under a fluorescence microscope is introduced using a cell‐friendly photoresist and spatially modulated light. Using this method, a single cell, regions in cell sheets, and a single focal adhesion complex within a cell are successfully detached. Furthermore, different subcellular regions of migrating cells are detached and changes in cell polarity and migration direction are quantitatively analyzed. This method will be useful for many applications in cell detachment, in particular when subcellular resolution is required.

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Programmed subcellular release for studying the dynamics of cell detachment

Cited by 42 publications

References 16 publications

Controlled two-photon photodegradation of PEG hydrogels to study and manipulate subcellular interactions on soft materials

Controlled two-photon photodegradation of PEG hydrogels to study and manipulate subcellular interactions on soft materials

Harvesting of Living Cell Sheets by the Dynamic Generation of Diffractive Photothermal Pattern on PEDOT

In Situ Subcellular Detachment of Cells Using a Cell‐Friendly Photoresist and Spatially Modulated Light

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