Journal: International Journal of Nanomedicine

Article Title: Human serum albumin-based probes for molecular targeting of macrophage scavenger receptors

doi: 10.2147/IJN.S197990

Figure Lengend Snippet: Evaluation of Mal-HSA-FITC uptake by different cell types. Fluorescence microscopy images of Mal-HSA-FITC probes (green) incubated for 3 h with ( A ) THP-1 macrophages, ( C ) HUVECs (EC), and ( D ) HCtSMCs (SMC). ( B ) Mal-HSA-FITC uptake by different cell types plotted against time. The results represent data from technical triplicates (9 images/well and timepoint) and are presented as mean and SEM. Data show significantly increased uptake of modified HSA probes by THP-1 cells compared with HUVEC (**** P <0.0001) and HCtSMCs ( #### P<0.0001) at 2.5 h and 3 h. None of the cell types recognized HSA-FITC (data not shown) (repeated measures two-way ANOVA (factors: cell type and time) and Tukey’s multiple comparison test). Abbreviations: HSA, human serum albumin; GCU, green calibration unit.

Article Snippet: Human carotid smooth muscle cells (HCtSMCs) (Cell Applications, San Diego, CA, USA) were cultured up to passage 7 in human smooth muscle cell media (Cell Applications).

Techniques: Fluorescence, Microscopy, Incubation, Modification, Comparison

Journal: bioRxiv

Article Title: A Mesenchymal Cell Niche in Skin for Acute Myeloid Leukemia

doi: 10.1101/2022.05.20.491183

Figure Lengend Snippet: ( A ) A representative FACS profile showing FACS sorting/analysis of the Ebf2 + and Ebf2 − cells in dorsal skin. The cells were first gated within non-hematopoietic (CD45 − TER119 − ) and non-endothelial (CD31 − ) live (PI − ) stromal cells. Then, these cells lacking expression of CD44 were further analyzed for their expression of SCA1, PDGFRa/CD140a (PαS) and CD51. The numbers in the panel are the mean frequencies. ( B-C ) The Ebf2 + cell frequency within total PI − ( B ) or PI − CD45 − TER119 − CD31 − stromal cells ( C ) in dorsal skin. ( D ) The fractions of PαS cells within the Ebf2 + and Ebf2 − stromal cells. Each dot in B-D represents data from a single mouse in 3-6 experiments with the horizontal line as a mean value. ( E ) CFU-Fs in the Ebf2 + and Ebf2 − stromal cells. ( F ) CFU-Fs were exclusively found in the Ebf2 + SCA1 + cell fraction. Data in E-F are from 3 independent experiments and each dot represents replicate assays from 2-3 mice in each experiment. The horizontal line represents mean value. Wilcoxon matched-signed pair rank test was used for statistical analysis. (G-J) Single-cell analysis of CFU-Fs and lineage differentiation from the FACS-sorted Ebf2 + ( G-H ) and Ebf2 − PαS ( I-J ) stromal cells. The CFU-F frequencies ( G, I ) were determined by limiting dilution at a density of 1, 2, 5, 10 cells per well in a 96-well plate and the frequency of the single cells with bi-lineage plasticity ( H, J ) were assessed by multilineage differentiation potential of single CFU-Fs derived from the cells. ( K ) Representative images of the osteogenic and adipogenic differentiation from single CFU-F clones derived from Ebf2 + and Ebf2 − PαS cells. ( L ) Population-doubling time (PDT) of randomly selected CFU-Fs derived from single Ebf2 + and Ebf2 − PαS cells. Each line represents the growth kinetics of a single clone. See also in Figure S1.

Article Snippet: For osteogenic differentiation, cells were cultured with complete osteogenic medium mixed by human/mouse StemXVivo osteogenic / adipogenic base medium (CCM007, R&D Systems) and mouse StemXVivo ostegenic supplement (CCM009; R&D Systems) under normoxic condition for 14-21 days.

Techniques: Expressing, Single-cell Analysis, Derivative Assay, Clone Assay

Journal: Biological Chemistry

Article Title: Peritumoral administration of GPI-anchored TIMP-1 inhibits colon carcinoma growth in Rag-2 γ chain-deficient mice

doi: 10.1515/bc.2009.098

Figure Lengend Snippet: Figure 2 The GPI-anchored TIMP-1 suppresses proliferation of SW480 cells and inhibits tumor invasion. The effect of cell surface engineering with TIMP-1-GPI on SW480 invasion through the Matrigel basement membrane model was assessed. Optimal invasion of the SW480 cells in response to 10% FCS, 4 ng/ml EGF, or 4 ng/ml VEGF were set to ‘zero’, and the 100% inhibition value was set to the migration level observed with untreated SW480 cells (Djafarzadeh et al., 2004). 5=103 SW480 cells per 100 ml medium were cultured in 96-well microtiter plates for 24 h under standard conditions to yield firmly attached and stably growing cells (Djafarzadeh et al., 2006). The SW480 cells were then pretreated with 2, 4, 6, 8, 10, 12, and 14 ng/ml of TIMP-1-GPI or 14 ng/ml of rhTIMP-1 as described previously (Djafarzadeh et al., 2006). After 1 h the cells were washed. (A) The effect of increasing levels of TIMP-1-GPI or rhTIMP-1 control protein on the proliferation of SW480 was measured using an MTT assay (Djafarzadeh et al., 2006). MTT was added after 24 h, 48 h, or 72 h as indicated. (B) The effect on the ability of SW480 to invade Matrigel was measured using a modified Boyden chamber as described earlier (Djafarzadeh et al., 2004, 2006).

Article Snippet: The effect of TIMP-1-GPI vs. rhTIMP-1 treatment on the ability of SW480 cells to invade Matrigel was measured using induced migration to fetal calf serum (FCS) (Biocrom, Berlin, Germany), vascular endothelial growth factor (VEGF) (R&D systems, Minneapolis, USA), and EGF (Biochrom, Berlin, Germany) using a modified Boyden chamber (Djafarzadeh et al., 2004, 2006).

Techniques: Membrane, Inhibition, Migration, Cell Culture, Stable Transfection, Control, MTT Assay, Modification