Microfabrication of high-resolution porous membranes for cell culture

Kim, Monica Y.; Li, David Jiang; Pham, Long K.; Wong, Brandon G.; Hui, Elliot E.

doi:10.1016/j.memsci.2013.11.034

Cited by 67 publications

(67 citation statements)

References 20 publications

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“…Polymeric track-etched membranes are available with a variety of pore sizes including sub-micron pores that prevent transmigration, but they are typically 10 μm in thickness and hinder physical contact between co-cultured cells on opposite faces of the membrane [17]. Additionally, track-etched membranes suffer from significant scattering in light microscopy [18], limiting live-imaging and requiring users to cut out and mount membranes post-fixation [19]. Research membranes have been fabricated using novel techniques, but most are still thicker than physiological barriers such as the basement membrane [10,18], which is just several hundred nanometers in thickness [20].…”

Section: Introductionmentioning

confidence: 99%

Ultrathin transparent membranes for cellular barrier and co-culture models

et al. 2017

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Typical in vitro barrier and co-culture models rely upon thick semi-permeable polymeric membranes that physically separate two compartments. Polymeric track-etched membranes, while permeable to small molecules, are far from physiological with respect to physical interactions with co-cultured cells and are not compatible with high-resolution imaging due to light scattering and autofluorescence. Here we report on an optically transparent ultrathin membrane with porosity exceeding 20%. We optimize deposition and annealing conditions to create a tensile and robust porous silicon dioxide membrane that is comparable in thickness to the vascular basement membrane (100–300 nm). We demonstrate that human umbilical vein endothelial cells (HUVECs) spread and proliferate on these membranes similarly to control substrates. Additionally, HUVECs are able to transfer cytoplasmic cargo to adipose-derived stem cells when they are co-cultured on opposite sides of the membrane, demonstrating its thickness supports physiologically relevant cellular interactions. Lastly, we confirm that these porous glass membranes are compatible with lift-off processes yielding membrane sheets with an active area of many square centimeters. We believe that these membranes will enable new in vitro barrier and co-culture models while offering dramatically improved visualization compared to conventional alternatives.

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Section: Introductionmentioning

confidence: 99%

Ultrathin transparent membranes for cellular barrier and co-culture models

et al. 2017

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“…However, these effects are not likely seen to the same degree on track-etched polymeric membranes due to highly variable pore density and greater pore spacing. 36,37 …”

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confidence: 99%

Membrane Pore Spacing Can Modulate Endothelial Cell–Substrate and Cell–Cell Interactions

Casillo

Peredo

Perry

et al. 2017

ACS Biomater. Sci. Eng.

133

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Mechanical cues and substrate interaction affect the manner in which cells adhere, spread, migrate and form tissues. With increased interest in tissue-on-a-chip and co-culture systems utilizing porous membranes, it is important to understand the role of disrupted surfaces on cellular behavior. Using a transparent glass membrane with defined pore geometries, we investigated endothelial fibronectin fibrillogenesis and formation of focal adhesions as well as development of intercellular junctions. Cells formed fewer focal adhesions and had shorter fibronectin fibrils on porous membranes compared to non-porous controls, which was similar to cell behavior on continuous soft substrates with Young’s moduli seven orders of magnitude lower than glass. Additionally, porous membranes promoted enhanced cell-cell interactions as evidenced by earlier formation of tight junctions. These findings suggest that porous membranes with discontinuous surfaces promote reduced cell-matrix interactions similarly to soft substrates and may enhance tissue and barrier formation.

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“…The chambers are divided by the 1002F resin-based membrane, and sealed by plasma bonding. The 1002F membrane is biocompatible, has a 6-μm thickness (DekTak3 surface profilometer) and 2-μm circular pores that allow soluble factor communication and cellular contact between cells cultured on the two sides 29. The patterned membrane fully spans the chamber to provide a compartmentalized region, suitable for cell co-culture assays (Fig.…”

Section: Resultsmentioning

confidence: 99%

Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium

Gnecco

Pensabene

et al. 2017

Ann Biomed Eng

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The endometrium is the inner lining of the uterus. Following specific cyclic hormonal stimulation, endometrial stromal fibroblasts (stroma) and vascular endothelial cells exhibit morphological and biochemical changes to support embryo implantation and regulate vascular function, respectively. Herein, we integrated a resin-based porous membrane in a dual chamber microfluidic device in polydimethylsiloxane that allows long term in vitro co-culture of human endometrial stromal and endothelial cells. This transparent, 2-μm porous membrane separates the two chambers, allows for the diffusion of small molecules and enables high resolution bright field and fluorescent imaging. Within our primary human co-culture model of stromal and endothelial cells, we simulated the temporal hormone changes occurring during an idealized 28-day menstrual cycle. We observed the successful differentiation of stroma into functional decidual cells, determined by morphology as well as biochemically as measured by increased production of prolactin. By controlling the microfluidic properties of the device, we additionally found that shear stress forces promoted cytoskeleton alignment and tight junction formation in the endothelial layer. Finally, we demonstrated that the endometrial perivascular stroma model was sustainable for up to 4 weeks, remained sensitive to steroids and is suitable for quantitative biochemical analysis. Future utilization of this device will allow the direct evaluation of paracrine and endocrine crosstalk between these two cell types as well as studies of immunological events associated with normal vs. disease-related endometrial microenvironments.Electronic supplementary materialThe online version of this article (doi:10.1007/s10439-017-1797-5) contains supplementary material, which is available to authorized users.

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Microfabrication of high-resolution porous membranes for cell culture

Cited by 67 publications

References 20 publications

Ultrathin transparent membranes for cellular barrier and co-culture models

Ultrathin transparent membranes for cellular barrier and co-culture models

Membrane Pore Spacing Can Modulate Endothelial Cell–Substrate and Cell–Cell Interactions

Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium

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