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Journal: Journal of Cellular Physiology
Article Title: Maternal Obesity Programs Adipogenic Commitment in Neonatal Mesenchymal Stem Cells: A Link to Redox‐Dependent FOXO1 Signaling
doi: 10.1002/jcp.70178
Figure Lengend Snippet: OB‐MSCs exhibit lower stemness, clonogenic capacity, and growth rate. (A) Wharton jelly‐derived mesenchymal stem cells were positive for surface markers CD73, CD90, and CD105, while negative for CD34, CD45, and CD11b in NW‐MSCs and O•B‐MSCs ( n = 3). The gating of cells is plotted as forward scatter versus side scatter for the cell population and forward scatter versus forward scatter for singlets (top). Representative graphs show surface markers of NW‐MSCs. (B) MSCs were cultured in induction media (osteogenic and myogenic) for 14 days, and in adipogenic induction medium for 21 days. MSCs were stained for lipids (Oil Red O), calcium (Alizarin Red), and α‐SMA (phalloidin) ( n = 3 NW‐MSCs and 3 OB‐MSCs). Representative images show induction of NW‐MSCs (Scale bars Oil Red O and Alizarin Red: 100 μm; Phalloidin: 400 μm). (C) MSCs from normal weight and obesity groups stained for Stage Specific Embryonic Antigen 4 (SSEA‐4) (scale bar: 100 μM). (D) OB‐MSCs showed lower gene expression levels of the stemness marker OCT3/4, compared to NW‐MSCs ( n = 5). (E) Representative images of colonies per well with magnification (left) and quantification of positive colonies at day 14 (right, n = 6 NW‐MSCs and 6 OB‐MSCs). (F) Growth curve of cell populations (left, n = 5 NW‐MSCs and 5 OB‐MSCs). Area under the curve (AUC) of NW‐MSCs and OB‐MSCs growth for 10 days (right). * p < 0.05 Mann−Whitney U test; ** p < 0.01 Tukey's range post hoc test.
Article Snippet: Surface markers were characterized according to the International Society of Cell Therapy criteria (Dominici et al. 2006) Briefly, cells were detached with trypsin and centrifuged at 2500 g for 3 min. After this, 1 × 10 6 cells were resuspended in 100 μL of FACS buffer (2% FCS in DPBS) in FACS tubes (Falcon, Corning) and incubated for 30 min in the dark with antibodies for CD73 (APC, 1:50, Miltenyi Biotec), CD90 (PE, 1:50, Miltenyi Biotec), CD105 (BV421, 1:20, BioLegend), CD34 (PE‐Dazzle 594, 1:20, Biolegend), CD45 (BV785, 1:20, BioLegend), and
Techniques: Derivative Assay, Cell Culture, Staining, Gene Expression, Marker, MANN-WHITNEY