primary human alveolar epithelial cells haecs  (PromoCell)

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: Concept of the 3D lung-on-a-chip model based on biomimetically microcurved culture membranes. (A) We approached a structure similar to a cut and flipped open alveolar sac as a cell-populated membrane with the microcurved shape, size, and also arrangement of its bioartificial alveoli mimicking the ones of the adult organ. Integrated in microfluidic chips/OoC devices where the microcurved membranes separate a top from a bottom compartment, they can be seeded by infusion with lung epithelial and microvascular endothelial cells on the top and bottom side of the membrane. The spatial cell distribution is then similar to the alveolar–capillary barrier. (B) Potential future applications of the model include 3D ALI culture (following submerged culture), modeling of disease and repair/regeneration, and toxicity and pharmaceutical efficacy testing (temporarily under submerged conditions or exposed to vapors or aerosols).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques: Membrane

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: Geometrical characterization of the (bottom) housing half of the chip from PDMS, and perfusion and leak test of the assembled OoC device. (A) Each housing half of the microfluidic chip contained one of the two central circular culture chambers with a diameter of 8 mm for receiving the hexagonal microwell array. This chamber was on either side connected to an inlet and an outlet channel with in each case a width of 500 μm and a length of 4 mm. At their lateral/peripheral ends across the culture chamber, the two channels in turn were connected to in each case one smaller chamber with a diameter of 1 mm located in two opposite corners of the chip. The height/depth of the microfluidic compartments was around 400 μm. (B) Cross-section of an assembled 3D lung-on-chip device (stitched image; housing halves that during cell culture host epithelial and endothelial cells are colored blue and pink/purple, respectively; scale bar represents 500 μm). (C) Assembled lung-on-chip device with its top and bottom chip compartment perfused through press-fitted tubing with water colored with green and blue (food) dye, respectively (scale bar represents 8 mm).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques: Cell Culture

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: Epithelialization of the microcurved membrane in the chip. HAECs cultured submerged under flow for 7 days and stained for cell nuclei and (A) F-actin, (B) tight junctions, (C) vimentin, and (D) CK8 (fluorescent microscopy images; nuclei not shown in the right halves of the images for better visibility of the individual stains; scale bars represent 100 μm).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques: Membrane, Cell Culture, Staining, Microscopy

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: ALI culture on the microcurved membrane in the chip. HAECs cultured at the ALI under perfusion for 14 days and stained for cell nuclei and (A) F-actin, (B) CK8, (C) aquaporin 5, and (D) pSPC (fluorescent microscopy images; scale bars represent 100 μm).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques: Membrane, Cell Culture, Staining, Microscopy

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: Thickness of the formed, curved alveolar epithelial layer. The thickness of the HAEC lining was measured (A) in two perpendicular cross-sections and there in each case in five different locations: at the horizontal center of the bottom of the microwell, at the left and right sidewall of the microwell directly under its convex rim, and at the left and right sidewall roughly halfway between, in each case perpendicular to the microwell wall. (B) Representative vertical and horizontal cross-sectional images of the epithelial layer (image planes “ x ” and “ y ”, and “ z 1 ” to “ z 3 ”, respectively; scale bars represent 50 μm). (C) Graph of the HAEC layer thickness as a function of the measurement location as stated in (A) ( n = 3).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques:

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: Lung epithelial and endothelial coculture on the microcurved membrane in the chip. (A) Top views of sections of the microcurved membrane with Calu-3 cells cultured for 11 days and stained for cell nuclei and tight junctions (fluorescent microscopy image; scale bars represent 100 μm). (B) Bottom views of sections of the same microcurved membrane with HLMVECs cultured for 11 days and stained for nuclei and CD31 (fluorescent microscopy image; scale bars represent 100 μm). (C) Cross-section of the microcurved membrane from (A) and (B) (scale bar represents 100 μm). (D) Graph of the count of Calu-3 cells ( n = 4) and HLMVECs per square microwell unit ( n = 3) (**** indicates a p -value smaller than 0.0001).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques: Membrane, Cell Culture, Staining, Microscopy

Journal: Scientific Reports

Article Title: K 2P 2.1 (TREK-1) potassium channel activation protects against hyperoxia-induced lung injury

doi: 10.1038/s41598-020-78886-y

Figure Lengend Snippet: TREK-1 activation with ML335 and BL1249 regulates cytokine secretion from primary human alveolar epithelial cells (HAEC): HO exposure increased secretion of IL-6, IP-10, CCL-2 and IL-10, and this effect was counteracted by ML335 or BL1249 ( A , B , C , F ). In contrast, TNF-α and MIP-1α levels were not affected by TREK-1 activation in room air- or HO-exposed HAECs ( D , E ). Data are represented as Box-Whisker plots with medians, 1st and 3rd quartiles, and max and min values; n = 4–8; ^compared to cells treated with a vehicle control and exposed to room air (no drugs), *compared to HO exposed cells; p ≤ 0.05.

Article Snippet: Primary Human Alveolar Epithelial Cells (HAEC) were purchased from ScienCell (#3200), cultured according to the company’s instructions, and used at a passage numbers < P5.

Techniques: Activation Assay, Whisker Assay, Control