Fabrication of ultrathin artificial basement membrane for epithelial cell culture

Peng

ACS Appl. Mater. Interfaces

et al. 2021

Self Cite

The development of in vitro neural networks depends to a large extent on the scaffold properties, including the scaffold stiffness, porosity, and dimensionality. Herein, we developed a method to generate interconnected neural clusters in a multiscale scaffold consisting of a honeycomb microframe covered on both sides with a monolayer of crosslinked gelatin nanofibers. Cortical neural precursor cells (NPCs) were firstly produced from human induced pluripotent stem cells (hiPSCs) and then loaded into the scaffold for a long period of differentiation toward cortical neural cells. As result, neurons and astrocytes self-organized in the scaffold to form clusters in each of the honeycomb compartments with remarkable inter-cluster connections. These cells highly expressed neuron-and astrocyte-specific proteins, including NF200, Tau, Synapsin I and GFAP, and showed spatially correlated neural activities. Two types of neural clusters, i.e., spheroid-like and hourglass-like clusters, were found, indicating the complexity of neural-scaffold interaction and the variability of three-dimensional neural organization. Furthermore, we incorporated a reconstituted basement membrane into the scaffold and performed coculture of the neural network with brain microvascular endothelial cells. As a proof of concept, an improved neurovascular unit model was tested showing large astrocytic endfeet on the backside of the endothelium.

Section: Discussionsupporting

confidence: 65%

Section: Fabrication Of Patterned Bilayer Nanofiber Scaffoldsmentioning

confidence: 99%

Generation of Interconnected Neural Clusters in Multiscale Scaffolds from Human-Induced Pluripotent Stem Cells

Peng

ACS Appl. Mater. Interfaces

et al. 2021

Self Cite

“…The MNF was first fabricated by electrospinning with a patterned honeycomb frame and then used as backbone for self‐assembling type IV‐collagen and laminin. [ 34 ] A porosity of ≈25% and a layer thickness of ≈200 nm could be estimated for a typical MNF. After the protein self‐assembling, MNF was converted to ABM with a comparable layer thickness but different bioactivity, stiffness, and permeability.…”

Section: Resultsmentioning

confidence: 99%

“…The in vivo like or artificial BM (ABM) was obtained by self‐assembling the two BM proteins with a monolayer of crosslinked gelatin nanofibers (MNF) and a handling honeycomb microframe. [ 34 ] Without BM proteins, the MNF could be used for both hiPSC differentiation of cardiomyocytes and neurons [ 35,36 ] and primary cell culture. [ 37–39 ] After BM protein self‐assembling, the MNF can be considered as backbone of the ABM.…”

Section: Introductionmentioning

confidence: 99%

Generation of Alveolar Epithelium Using Reconstituted Basement Membrane and hiPSC‐Derived Organoids

Rofaani

Adv Healthcare Materials

et al. 2022

Self Cite

In vitro modeling of alveolar epithelium needs to recapitulate features of both cellular and noncellular components of the lung tissues. Herein, a method is presented to generate alveolar epithelium by using human induced pluripotent stem cells (hiPSCs) and reconstituted or artificial basement membrane (ABM). The ABM is obtained by self‐assembling type IV collagen and laminin with a monolayer of crosslinked gelatin nanofibers as backbone and a patterned honeycomb microframe for handling. Alveolar organoids are obtained from hiPSCs and then dissociated into single cells. After replating the alveolar cells on the ABM and a short‐period incubation under submerged and air–liquid interface culture conditions, an alveolar epithelium is achieved, showing high‐level expressions of both alveolar cell‐specific proteins and characteristic tight junctions. Besides, endothelial cells derived from the same hiPSCs are cocultured on the backside of the epithelium, forming an air–blood barrier. The method is generic and can potentially be applied to other types of artificial epithelium and endothelium.

“…Newer second generation LOAC devices have recently been created replacing the PDMS material with stretchable membranes made of collagen and elastin to better emulate the three-dimensional alveolar structure [34]. Other groups have created improved artificial basement membranes to separate different cell types through a combination of lithography and vacuum assisted UV curing techniques by electrospun gelatin nanofibers [35]. To this end, we sought to evaluate a new material substrate (porous silicon) for structural membranes based on a bottom-up fabrication technique that allow for cell adherence along with exhibiting properties of biocompatibility and mechanical flexibility.…”

Section: Introductionmentioning

confidence: 99%

Novel Fabrication Approach of a Porous Silicon Biocompatible Membrane Evaluated within a Alveolar Coculture Model

Williams¹,

Wiens²,

Hamilton³

et al. 2021

Preprint

The use of conventional in vitro and preclinical animal models often fail to properly recapitulate the complex nature of human diseases and hamper the success of translational therapies in humans [1-3] Consequently, research has moved towards organ-on-chip technology to better mimic human tissue interfaces and organ functionality. Herein, we describe a novel approach for the fabrication of a biocompatible membrane made of porous silicon (PSi) for use in organ-on-chip technology that provides key advantages when modeling complex tissue interfaces seen in vivo. By combining well-established methods in the semiconductor industry with organ-on-chip technology, we have developed a novel way of producing thin (25 μm) freestanding PSi biocompatible membranes with both nano (~15.5 nm diameter pores) and macroporous (~0.5 μm diameter pores) structures. To validate the proposed novel membrane, we chose to recapitulate the dynamic environment of the alveolar blood gas exchange interface in alveolar co-culture. Viability assays and immunofluorescence imaging indicate that human pulmonary cells remain viable on the PSi membrane during long-term culture (14 days). Interestingly, it was observed that macrophages can significantly remodel and degrade the PSi membrane substrate in culture. This degradation will allow for more intimate physiological cellular contact between cells, mimicking a true blood-gas exchange interface as observed in vivo. Broadly, we believe that this novel PSi membrane may be used in more complex organ-on-chip and lab-on-chip model systems to accurately recapitulate human anatomy and physiology to provide further insight into human disease pathology and pre-clinical response to therapeutics.