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