Journal: Cells

Article Title: Extracellular Lactic Acidosis of the Tumor Microenvironment Drives Adipocyte-to-Myofibroblast Transition Fueling the Generation of Cancer-Associated Fibroblasts

doi: 10.3390/cells12060939

Figure Lengend Snippet: Quantitative real-time PCR analysis for the expression of the fibroblast activation/myofibroblast genes FAP ( A ), ACTA2 ( B ), COL1A1 ( C ), and COL1A2 ( D ) in normal human dermal fibroblasts treated for 24 h with conditioned media produced by adipocyte-committed adipose-derived stem cells (acADSCs) grown either under basal (pH 7.4) conditions (basal acADSC-cm) or under lactic acidosis (acidic acADSC-cm). The basal level of each gene expression was set to 1, and the other results are normalized to this value. 18S ribosomal RNA was used as the reference gene. Bars represent the mean ± SEM of three independent experiments ( n = 3 replicates each) from three cell lines. Values of p were determined by the unpaired Student’s t -test.

Article Snippet: Three lines of normal human subcutaneous adipose-derived stem cells (ADSCs) purchased from Lonza (catalog no. PT-5006; Lonza, Basel, Switzerland) were maintained in ADSC complete medium (ADSC Growth Medium BulletKit; catalog no. PT-4505; Lonza) at 37 °C in a 5% CO 2 incubator.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Activation Assay, Produced, Derivative Assay

Journal: Regenerative Therapy

Article Title: Increase of gremlin 2 with age in human adipose-derived stromal/stem cells and its inhibitory effect on adipogenesis

doi: 10.1016/j.reth.2019.09.002

Figure Lengend Snippet: GREM2 expression in young and old adipose tissues. Immunohistochemistry was performed on subcutaneous adipose tissues obtained from 36 subjects 12–97 years of age (A) Representative HE staining images of adipose tissues (upper panel) and immunofluorescence images against GREM2 (lower). Young, adipose tissue from the back of a 23-year-old women; old, from the lumbar region of a 69-year-old men. Bars = 100 ㎛ (B) The average numbers of cells stained with DAPI except mature adipocytes in immunofluorescence images of subcutaneous adipose tissue areas (200 μm × 200 μm) were plotted (C) Integrated fluorescent intensities of GREM2 were calculated for each area. The value for the young sample shown in (A) derived from a 23-year-old subject was set as 1, and the relative values of GREM2 integrated fluorescent intensities were plotted (D) GREM2 integrated fluorescent intensities adjusted by cell number were plotted. For each, Pearson's product–moment correlation analysis (a parametric method) was performed to assess the degree of relationship. ** p < 0.01.

Article Snippet: ASCs collected from adipose tissues and human adipose derived stem cells (ADSCs; from a Hispanic female 34 years of age; cell strain No. 01171; KURABO, Osaka, JPN) were cultured in complete medium (D/α medium), which consists of Dulbecco's modified Eagle's medium (DMEM, Invitrogen) and α minimum essential medium (α MEM, Invitrogen) with a 1:1 ratio, supplemented with 1% fetal bovine serum (Sigma–Aldrich, MO, USA), 1 × ITS-X (Invitrogen), 10 ng/mL basic FGF (PeproTech, NJ, USA), 0.4 μg/mL hydrocortisone, and 1% Antibiotic-Antimycotic (Gibco BRL, MD, USA) at 37 °C in a humidified atmosphere with 5% CO 2 .

Techniques: Expressing, Immunohistochemistry, Staining, Immunofluorescence, Derivative Assay