aim 3d cell culture chips  (AIM Biotech)

Journal: Small methods

Article Title: A Robust Method for Perfusable Microvascular Network Formation In Vitro

doi: 10.1002/smtd.202200143

Figure Lengend Snippet: Initial cell spatial distribution determines MVN morphology. a) Schematic illustration of non-flipped and flipped seeding method. EC-FB mixture was seeded in the microfluidic device, the device either stayed right side up (no flipping), or was flipped —three to five times. The cell mixture is more evenly distributed in all three dimensions in flipped devices, leading to narrower and more 3D distributed MVNs at day 7, compared to non-flipped method. Representative confocal images of ImHUVECs expressing GFP and MVNs seeding in non-flipped b) or flipped c) method at day 0 (left) and day 7 (right). d) Diameter distribution of microvessels made in non-flipped or flipped method. Mean and SD of each group are: non-flipped 63.27 ± 45.29 μm; flipped 37.17 ± 19.00 μm (n = 3 devices, 2 ROIs each).

Article Snippet: 3D cell culture chips (AIM Biotech) were utilized to generate in vitro MVNs.

Techniques: Expressing

Journal: Small methods

Article Title: A Robust Method for Perfusable Microvascular Network Formation In Vitro

doi: 10.1002/smtd.202200143

Figure Lengend Snippet: Two-step strategy for robustly generating perfusable MVNs of physiological geometry. a) Schematic diagram of two-step seeding method in the microfluidic device with microposts. All cells were suspended in hydrogel precursor (fibrinogen/thrombin). First, high density of ECs are seeded in the device. Second, the uncured gel is carefully aspirated, and the ECs remaining in the regions between microposts are referred to as outer layer ECs. Third, the ECs/stromal cells, such as FBs, are seeded into the central gel region. Microfluidic devices are flipped several times to improve vascular cell 3D distribution in the center. MVNs will form within 7 days. b) Schematic illustration and representative images of the traditional 1-step method of forming MVNs. 1) Non-flipped, higher EC (8 × 10 6 mL −1 ) with lower FB (1 × 10 6 mL −1 ) cell density tend to form perfusable but wider diameter MVNs. 2) Evenly distributed (flipped), lower EC (5 × 10 6 mL −1 ) with higher FB (1.5 × 10 6 mL −1 ) cell density form non-perfusable, narrower diameter MVNs. ImHUVECs expressing BFP and lung FBs were used. Green, ImHUVECs. Magenta, dextran (70 kDa). c) Schematic cartoon and representative images of the two-step method of generating perfusable MVNs with narrower diameters in the center. In this experiment, BFP-ImHUVECs (cyan, 10 × 10 6 mL −1 ) were used as outer layer ECs; GFP-ImHUVECs (green, 5 × 10 6 mL −1 ) with FBs (1.5 × 10 6 mL −1 ) were seeded in the center. Devices were flipped several times during hydrogel polymerization. The dotted square indicates an enlarged dextran image that demonstrates perfusibility of the narrow vessels. White arrows point to narrow microvessels. d) Diameter distribution of microvessels from MVNs made by one- or two-step seeding method. Mean and SD of each group are one-step: perfusable but wider vessels 74.17 ± 62.27 μm; one-step: narrow vessels but not perfusable 21.16 ± 12.95 μm; two-step: outer layer 80.60 ± 25.36 μm; two-step: center 27.46 ± 18.63 μm (n = 3 devices, 3 ROIs each). e) Statistical analysis of microvessel opening percentage (bars) and central perfusable MVN percentage (dots) in each ROI of MVNs made by one- or two-step seeding method. Bars represent mean ± SD. Two-tailed t -tests were performed for the statistical comparisons (n = 3 devices, 5 ROIs each).

Article Snippet: 3D cell culture chips (AIM Biotech) were utilized to generate in vitro MVNs.

Techniques: Expressing, Two Tailed Test

Journal: Frontiers in Molecular Biosciences

Article Title: Boosting Natural Killer Cell Therapies in Glioblastoma Multiforme Using Supramolecular Cationic Inhibitors of Heat Shock Protein 90

doi: 10.3389/fmolb.2021.754443

Figure Lengend Snippet: SCI-101 increases NK92-MI tumor infiltration and cell killing in a 3D tumor spheroid model of GBM. (A) Microfluidic device and experimental design. GBM tumor spheroids are embedded in a collagen matrix and treated with SCI-101 as described in Materials and Methods . CFSE-labeled NK92-MI cells are then introduced to the media ports. Functional and quantitative readouts include infiltration capability of NK92-MI cells and tumor viability and growth. (B) Representative fluorescent microscopy overlaying brightfield image of NK92-MI cells (green) in U-251 tumor spheroids. NK cells infiltrate the tumor collagen chamber as indicated by a dashed arrow. Scale bar = 100 μM. The right panel quantifies the relative fluorescence of the CFSE signal in tumor spheroids, as described in Materials and Methods . *p<0.05 by the unpaired t-test. (C) Representative fluorescent microscope image of U-251 spheroids +/- SCI-101 (10 μM) and NK92-MI (E:T 5:1). (D) (left panel) Histogram quantifying normalized NK cell killing in vehicle controls and SCI-101–treated spheroids with 5 μM and 10 μM. (Right panel) NK cell killing in vehicles and SCI-101–treated (10 μM) U-87 spheroids. Values are determined by H/PI image analysis as described in Materials and Methods . *p<0.05 by the unpaired t-test. (E) Histogram quantifying the live cell area of U-251 spheroids +/- SCI-101 (5 μM) and NK92-MI (E:T 5:1). *p<0.05 by the unpaired t-test.

Article Snippet: The spheroid/collagen mixture was then injected into the center gel channel of an AIM 3D microfluidic cell culture chip (AIM Biotech) followed by 35 min incubation allowing collagen to polymerize.

Techniques: Labeling, Functional Assay, Microscopy, Fluorescence