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

Journal: Cells

Article Title: Generation of Functional Vascular Endothelial Cells and Pericytes from Keratinocyte Derived Human Induced Pluripotent Stem Cells

doi: 10.3390/cells10010074

Figure Lengend Snippet: Co-culture of human induced pluripotent stem cells (hiPSC)-derived endothelial cells (ECs) and pericytes. ( a ). Co-cultured hiPSC-derived ECs and pericytes form blood vessel-like structures in vitro under 2D culturing conditions. Fluorescent imaging shows blood vessel-like structures that contain ECs (positively stained for CD31) and pericytes (positively stained for Platelet Derived Growth Factor Receptor β (PDGFR-β)); scale bars: 100 μm. ( b ). Co-cultured hiPSC-derived ECs and pericytes form a 3D microvascular network in the chip. Live-cell imaging, both ECs, and PCs express mCherry for visualization; scale bars: 1 mm ( c ). Fluorescent imaging shows the 3D microvascular network in the chip, ECs and PCs express mCherry, ECs are positively stained for CD31. White arrows show the colocalization of pericytes with ECs; scale bars: 100 μm.

Article Snippet: In the project, 3D Cell Culture Chips (AIM Biotech, Nucleos, Singapore, Cat No: DAX01-1PAK) were used.

Techniques: Co-Culture Assay, Derivative Assay, Cell Culture, In Vitro, Imaging, Staining, Live Cell Imaging