Patch method for culture of primary hippocampal neurons

Tang, Yadong; Severino, Francesco Paolo Ulloa; Iseppon, Federico; Torre, Vincent; Chen, Yong

doi:10.1016/j.mee.2017.01.012

Cited by 17 publications

(24 citation statements)

References 19 publications

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“…The result is that the cells appear to be less rounded after the strain step in 3-day cultures which is evident from a decrease in roundness parameter of the cells. These results corroborate previous observations of good adhesion between gelatin nanofibers and other cells such as human pluripotent stem cells, motor neuron cells, primary hippocampal neurons and cardiomyocytes (Liu et al, 2014;Tang, Liu, Li, Yu, Severino, et al, 2016;Tang et al, 2017), and extends the applicability of gelatin nanofiber monolayers to alveolar cells.…”

Section: Figure 8(c)supporting

confidence: 91%

“…The use of gelatin is for its biocompatibility and its derivation from collagen which is the most abundant protein in the basement membrane (Dunsmore & Rannels, 1996;Maina & West, 2005;Townsley, 2012). The resulting monolayers have a high porosity which is desirable for a minimal exogeneous material contact with the cells, and a maximal exposure to the culture medium (Liu et al, 2014;Tang, Liu, Li, Yu, Severino, et al, 2016;Tang, Ulloa Severino, Iseppon, Torre, & Chen, 2017). The suspended monolayers have a thickness less than 1 µm and a relatively low elastic modulus affording their deformability at the airliquid interface.…”

Section: Introductionmentioning

confidence: 99%

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Alveolar mimics with periodic strain and its effect on the cell layer formation

Radiom

Peng‐Wang

et al. 2020

Biotech & Bioengineering

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We report on the development of a new model of alveolar air-tissue interface on a chip. The model consists of an array of suspended hexagonal monolayers of gelatin nanofibers supported by microframes and a microfluidic device for the patch integration. The suspended monolayers are deformed to a central displacement of 40 -80 µm at the air-liquid interface by application of air pressure in the range of 200 -1000 Pa. With respect to the diameter of the monolayers that is 500 µm, this displacement corresponds to a linear strain of 2 -10% in agreement with the physiological strain range in the lung alveoli. The culture of A549 cells on the monolayers for an incubation time 1 -3 days showed viability in the model. We exerted a periodic strain of 5% at a frequency of 0.2 Hz during 1 hour to the cells. We found that the cells were strongly coupled to the nanofibers, but the strain reduced the coupling and induced remodeling of the actin cytoskeleton, which led to a better tissue formation. Our model can serve as a versatile tool in lung investigations such as in inhalation toxicology and therapy.

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Section: Figure 8(c)supporting

confidence: 91%

Section: Introductionmentioning

confidence: 99%

Alveolar mimics with periodic strain and its effect on the cell layer formation

Radiom

Peng‐Wang

et al. 2020

Biotech & Bioengineering

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“…Previously, culture patches without collagen IV-laminin treatment could be used as substrates for differentiation of human induced pluripotent stem cells toward cardiomyocytes [18] and neurons [21] as well as primary neuron cells [22] and fibroblasts [23]. In such cases, collagen IVlaminin treatment would not be necessary since these cells were not epithelial.…”

Section: Resultsmentioning

confidence: 99%

Fabrication of ultrathin artificial basement membrane for epithelial cell culture

Rofaani

Peng

Wang

et al. 2020

Microelectronic Engineering

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Basement membranes are essential for epithelial and endothelial tissue organization. To mimic them, we developed a fabrication method to produce ultrathin membrane of collagen IV and laminin. A honeycomb microframe of thickness 50 µm and compartment-size 400 µm was firstly patterned in polyethylene glycol diacrylate (PEGDA) by using lithography and vacuum assisted UV curing techniques. Then, a monolayer of gelatin nanofibers was electrospun and crosslinked on the microframe to form a secondary structure, i.e. a fiber mesh with much smaller pore sizes, in each of the honeycomb compartments. Finally, a collagen IV-laminin gel layer was deposited and dehydrated, leading to the formation of an ultrathin membrane over a large area with a nanofiber backbone. Such an artificial basement membrane is mechanically stable and fully biocompatible. It is also semi-permeable which slowed down considerably the diffusion of large size molecules. More importantly, it could be used to improve the monolayer formation of epithelial cells. Thus, this culture device recapitulates the biological basement membrane, permits the epithelial tissue formation, and allows mimicking a more sophisticated cellular microenvironment.

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“…Our cage device is in the form of a patch with honeycomb frame sandwiched by a monolayer of agarose coated nanofibers and a mesh of relatively large openings. Both frame and mesh were defined by UV-lithography and soft-lithography and the nanofibers were produced by electrospinning, in a similar way of culture patch fabrication [19][20][21]. Single tumor cells could be dispersed through the mesh holes into each of the honeycomb compartments.…”

Section: Introductionmentioning

confidence: 99%

Fabrication of micro-cages and caged tumor spheroids for microfluidic chip-based assays

Huang

Rofaani

et al. 2020

Microelectronic Engineering

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We developed a simple method to fabricate micro-cages and caged tumor spheroids for microfluidic chip-based assays. The micro-cage device consists of an array of honeycomb compartments with a monolayer of cross-linked and agarose-coated gelatin nanofibers at the bottom and a mesh of 200 μm hole-size on the top. U87-MG single cells were dispersed through the mesh and resulted tumor spheroids confined in each of the cage compartment after incubation. As expected, the tumor spheroids are one-by-one distributed in each of the compartment with the same size and they grew inside the compartments. The final size of the spheroid was limited by both diffusion and confinement. If the height of the cage is small, the nanofiber layer underneath tumors could be deflected due to mechanic stress of growing tumors. If the height of the cage is large, tumors grew freely without stress but their size was limited by diffusion. In both cases, tumors tended to remain in spherical shape. To illustrate the robustness of the approach, the tumor caged device was reversibly integrated into a microfluidic chip for drug test. Our results show that under tangent flow conditions, combretastatin A-4 had a clear effect on tumor disassembling.

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Patch method for culture of primary hippocampal neurons

Cited by 17 publications

References 19 publications

Alveolar mimics with periodic strain and its effect on the cell layer formation

Alveolar mimics with periodic strain and its effect on the cell layer formation

Fabrication of ultrathin artificial basement membrane for epithelial cell culture

Fabrication of micro-cages and caged tumor spheroids for microfluidic chip-based assays

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