A Cell Culture System for the Structure and Hydrogel Properties of Basement Membranes: Application to Capillary Walls

Bruggeman, Leslie A.; Doan, Ryan P.; Loftis, Jacqueline; Darr, Adnan; Calabro, Anthony

doi:10.1007/s12195-012-0221-3

Cited by 15 publications

(13 citation statements)

References 32 publications

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“…Very few models have been developed to incorporate microvascular tissue to mimic reabsorptive barriers in vitro [ 33 , 34 ] . Transwells provide either a non-contact orientation with a 1 mm distance between the two cell types, or cells on both sides of the Transwell insert membrane to establish an approximation of a close-contact orientation of the two cell types[ 35 , 36 ].…”

Section: Resultsmentioning

confidence: 99%

A microfluidic renal proximal tubule with active reabsorptive function

et al. 2017

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In the kidney, the renal proximal tubule (PT) reabsorbs solutes into the peritubular capillaries through active transport. Here, we replicate this reabsorptive function in vitro by engineering a microfluidic PT. The microfluidic PT architecture comprises a porous membrane with user-defined submicron surface topography separating two microchannels representing a PT filtrate lumen and a peritubular capillary lumen. Human PT epithelial cells and microvascular endothelial cells in respective microchannels created a PT-like reabsorptive barrier. Co-culturing epithelial and endothelial cells in the microfluidic architecture enhanced viability, metabolic activity, and compactness of the epithelial layer. The resulting tissue expressed tight junctions, kidney-specific morphology, and polarized expression of kidney markers. The microfluidic PT actively performed sodium-coupled glucose transport, which could be modulated by administration of a sodium-transport inhibiting drug. The microfluidic PT reproduces human physiology at the cellular and tissue levels, and measurable tissue function which can quantify kidney pharmaceutical efficacy and toxicity.

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Section: Resultsmentioning

confidence: 99%

A microfluidic renal proximal tubule with active reabsorptive function

et al. 2017

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“…Few researchers have combined the Boyden chamber approach with collagen by eliminating the nonphysiological stiff membrane and suspending collagen membrane between side supports, thus creating a biomimetic culture platform [13,14,15,16]. For example, a casting method was used to create a thin layer of collagen between supports to mimic the glomerulas interface in kidneys with juxtaposition of podocytes and endothelial cells on either sides of the collagen film [15]. Another group employed a similar floating thin fibrillar collagen gel layer supported by rigid nylon support to investigate how physical boundaries interrupt the cell-collagen interactions based on the strain-stiffening nature of collagen [16].…”

Section: Introductionmentioning

confidence: 99%

A Novel Suspended Hydrogel Membrane Platform for Cell Culture

Chen

Yang

Yan

et al. 2015

Journal of Nanotechnology in Engineering and Medicine

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Current cell-culture is largely performed on synthetic two-dimensional (2D) petri dishes or permeable supports such as Boyden chambers, mostly because of their ease of use and established protocols. It is generally accepted that modern cell biology research requires new physiologically relevant three-dimensional (3D) cell culture platform to mimic in vivo cell responses. To that end, we report the design and development of a suspended hydrogel membrane (ShyM) platform using gelatin methacrylate (GelMA) hydrogel. ShyM thickness (0.25–1 mm) and mechanical properties (10–70 kPa) can be varied by controlling the size of the supporting grid and concentration of GelMA prepolymer, respectively. GelMA ShyMs, with dual media exposure, were found to be compatible with both the cell-seeding and the cell-encapsulation approach as tested using murine 10T1/2 cells and demonstrated higher cellular spreading and proliferation as compared to flat GelMA unsuspended control. The utility of ShyM was also demonstrated using a case-study of invasion of cancer cells. ShyMs, similar to Boyden chambers, are compatible with standard well-plates designs and can be printed using commonly available 3D printers. In the future, ShyM can be potentially extended to variety of photosensitive hydrogels and cell types, to develop new in vitro assays to investigate complex cell–cell and cell–extracellular matrix (ECM) interactions.

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“…The lamination of laser cut PE sheets (with pressure sensitive adhesive on one face) is compatible with commercially available micro and nanoporous membranes that have been optimized to provide low auto-fluorescence, high optical clarity, and defined pore size distributions. Materials such as ultrathin nano-membranes [ 47 , 48 ] collagen films [ 49 ], hydrogel sheets [ 50 ], patterned substrates [ 51 , 52 ], commercial EMC products [ 53 ], and layered composite biomaterials [ 54 ] can also be included to further tune the culture interface to reflect the physiology of the tissue.…”

Section: Discussionmentioning

confidence: 99%

A Reversibly Sealed, Easy Access, Modular (SEAM) Microfluidic Architecture to Establish In Vitro Tissue Interfaces

Abhyankar¹,

Wu²,

Koh³

et al. 2016

PLoS ONE

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Microfluidic barrier tissue models have emerged as advanced in vitro tools to explore interactions with external stimuli such as drug candidates, pathogens, or toxins. However, the procedures required to establish and maintain these systems can be challenging to implement for end users, particularly those without significant in-house engineering expertise. Here we present a module-based approach that provides an easy-to-use workflow to establish, maintain, and analyze microscale tissue constructs. Our approach begins with a removable culture insert that is magnetically coupled, decoupled, and transferred between standalone, prefabricated microfluidic modules for simplified cell seeding, culture, and downstream analysis. The modular approach allows several options for perfusion including standard syringe pumps or integration with a self-contained gravity-fed module for simple cell maintenance. As proof of concept, we establish a culture of primary human microvascular endothelial cells (HMVEC) and report combined surface protein imaging and gene expression after controlled apical stimulation with the bacterial endotoxin lipopolysaccharide (LPS). We also demonstrate the feasibility of incorporating hydrated biomaterial interfaces into the microfluidic architecture by integrating an ultra-thin (< 1 μm), self-assembled hyaluronic acid/peptide amphiphile culture membrane with brain-specific Young’s modulus (~ 1kPa). To highlight the importance of including biomimetic interfaces into microscale models we report multi-tiered readouts from primary rat cortical cells cultured on the self-assembled membrane and compare a panel of mRNA targets with primary brain tissue signatures. We anticipate that the modular approach and simplified operational workflows presented here will enable a wide range of research groups to incorporate microfluidic barrier tissue models into their work.

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A Cell Culture System for the Structure and Hydrogel Properties of Basement Membranes: Application to Capillary Walls

Cited by 15 publications

References 32 publications

A microfluidic renal proximal tubule with active reabsorptive function

A microfluidic renal proximal tubule with active reabsorptive function

A Novel Suspended Hydrogel Membrane Platform for Cell Culture

A Reversibly Sealed, Easy Access, Modular (SEAM) Microfluidic Architecture to Establish In Vitro Tissue Interfaces

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