Journal: Integrative Cancer Therapies

Article Title: Differential Effects of Wedelia chinensis on Human Glioblastoma Multiforme Cells

doi: 10.1177/15347354211000119

Figure Lengend Snippet: Effects of Wedelia chinensis extract (WCE) on the viability of GBM8401, U-87MG, and human brain astrocyte (HBA) cells. Relative cell survival of GBM8401, U-87MG, and HBA cells after treatment with various concentrations of WCE for (A) 24 and (B) 48 hours. Similar results were observed in 3 repeated experiments. The numbers 1, 2, and 3 indicate a significant difference ( P < .05) compared with control (0 μg/mL), HBA, and GBM8401 cells, respectively.

Article Snippet: Human brain astrocyte (HBA) cells and human malignant glioma cell lines, GBM8401 and U-87MG, were purchased from ScienCell (CA, USA) and ATCC (Rockville, MD, USA), respectively.

Techniques: Control

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Occludin regulates glucose uptake and ATP production in pericytes by influencing AMP-activated protein kinase activity

doi: 10.1177/0271678X17720816

Figure Lengend Snippet: Pericytes share glucose with astrocytes in an occludin-modulated manner. (a) Confocal microscopy of a representative live pericyte-astrocyte co-culture 30 min after plating. Pericytes were pre-loaded with 2-NBDG (green; e.g. blue thick arrows) and astrocytes with violet-BMQC (cell mask, red) but not with 2-NBDG. Colocalization of both signals (yellow-orange; e.g. white thin arrows) indicated that astrocytes had received 2-NBDG. Intensity coefficient (Ic) represents the whole 2-NBDG fluorescence intensity normalized to the surface it occupies, regardless of cell type. A larger Ic means more 2-NBDG was introduced into the system (taken up by pericytes). Transfer coefficient (Tc) represents the fraction of astrocytic surface occupied by 2-NBDG normalized against the Intensity coefficient. A larger Tc implicates more 2-NBDG was distributed across all possible astrocytes, and represents greater transferred volumes. Data correspond to the quantitation of the images shown. They are representative of three separate experiments. (b) Distribution of 2-NBDG in the same co-culture shown in (a), 24 h after plating. Pericytes are devoid of any stain (e.g. white thin arrows). 2-NBDG signal (green) colocalizing with astrocytes is seen as cyan/white (e.g. yellow thick arrows). (c) Similar co-culture as in (a) recorded 30 min post-plating; however, pericytes were treated with negative-control siRNA (SCR) before being loaded with 2-NBDG. (d) Distribution of 2-NBDG (green/cyan) in the same co-culture shown in (c), 24 h post-plating. (e) Similar co-culture as in (a) and (c), recorded 30 min post-plating; however, pericytes were treated with anti-occludin siRNA (OCC−) before being loaded with 2-NBDG. (f) Distribution of 2-NBDG (green/cyan) in the same co-culture shown in (e), 24 h post-plating. All images are representative of three separate experiments. (g) Average intensity (Ic) and H) Transfer (Tc) coefficients depicting transcellular glucose transport between pericytes and astrocytes as shown in (a) to (c). n = 3, p vs. WT.

Article Snippet: Cell culture Primary human brain capillary pericytes and astrocytes (ScienCell, Carlsbad, CA, USA) were cultured in 5% CO 2 at 37°C in pericyte or astrocyte growth medium (ScienCell), following standard cell culture procedures, and used between passages 2 and 7.

Techniques: Confocal Microscopy, Co-Culture Assay, Fluorescence, Quantitation Assay, Staining, Negative Control

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Occludin regulates glucose uptake and ATP production in pericytes by influencing AMP-activated protein kinase activity

doi: 10.1177/0271678X17720816

Figure Lengend Snippet: Pericytes share mitochondria with astrocytes in an occludin-mediated manner. (a) Live co-culture, 24 h post-plating, of astrocytes labeled with violet-BMQC (blue) and mitochondria-stained (TMRE in green) pericytes treated with anti-occludin siRNA (OCC−), negative-control siRNA (SCR), or non-treated (wild-type, WT). Thin white arrows exemplify TMRE-stained pericytes, while thick yellow arrows show pericyte mitochondria in the body of astrocytes (cyan signal). (b) Quantitation of TMRE intensities in astrocytes and pericytes in the same co-cultures shown in (a). Average ± SEM, n = 25 collected from three experiments, p vs. SCR. Only significant values are shown (c) Astrocytes treated with vehicle (Veh) or with endosulfan sulfate (ES, 4 h) to block their energetic metabolism. Middle image, surviving astrocytes exhibiting widened bodies and gross morphological alterations are exemplified by thin white arrows. Yellow arrowheads point to astrocytes that still retain their normal morphology. Right image: astrocytes treated with ES; however, isolated murine live brain capillaries (D shows a single brain capillary) were added to their growth medium 2 h post-treatment, and incubated for two additional hours. Note markedly improved astrocyte morphology. (e) Not-labeled human astrocytes cultured with murine live brain capillaries pre-labeled with TMRE (red) and 2-NBDG (green) for 2 h. TMRE and 2-NBDG transferred from microvessels to astrocytes (arrows) indicate transfer of mitochondria and glucose, respectively. (f) Similar TMRE and 2-NBDG transfer (arrows) in microvessel-rescued/ES-treated astrocytes after incubation with TMRE and 2-NBDG-labeled murine live brain capillaries.

Article Snippet: Cell culture Primary human brain capillary pericytes and astrocytes (ScienCell, Carlsbad, CA, USA) were cultured in 5% CO 2 at 37°C in pericyte or astrocyte growth medium (ScienCell), following standard cell culture procedures, and used between passages 2 and 7.

Techniques: Co-Culture Assay, Labeling, Staining, Negative Control, Quantitation Assay, Blocking Assay, Isolation, Incubation, Cell Culture

Journal: Journal of Cellular and Molecular Medicine

Article Title: Escitalopram oxalate induces apoptosis in U‐87 MG cells and autophagy in GBM 8401 cells

doi: 10.1111/jcmm.13372

Figure Lengend Snippet: The effects of escitalopram oxalate on the viability, motility and invasive abilities of U‐87 MG and human brain astrocyte ( HBA ) cells. Relative cell survival of U‐87 MG and HBA was detected after treatment with different concentrations of escitalopram oxalate for ( A ) 24 and ( B ) 48 hrs. ( C ) Wound‐healing assay and ( D ) transwell migration assay were performed in U‐87 MG cells treated with different concentrations of escitalopram oxalate for 24 hrs. Similar results were observed in three repeated experiments, and * indicates the significant difference, P < 0.05.

Article Snippet: Human brain astrocyte cells (HBA) and human malignant gliomas cell lines, GBM8401, U‐87MG and Hs683, were purchased from ScienCell (CA, USA) and ATCC (Rockville, MD, USA), respectively.

Techniques: Wound Healing Assay, Transwell Migration Assay