human ascs (hasc)  (Wolters Kluwer Health)

Journal: Lab on a Chip

Article Title: Adipose microtissue-on-chip: a 3D cell culture platform for differentiation, stimulation, and proteomic analysis of human adipocytes

doi: 10.1039/d2lc00245k

Figure Lengend Snippet: Parallel formation and cultivation of human adipose tissue-derived adult stem cells in 3D on-chip. A) Timeline of hASC self-aggregation in the cell culture chamber. Scale bar: 250 μm. B) Intrachip variability of the equivalent diameter of the 3D cell cultures. n = 6 biological repeats. C) Interchip variability of the equivalent diameter of formed hASC 3D cell cultures. n = 96 technical replicates per chip, all comparisons exhibited a p < 0.05 (one-way ANOVA) except for n.s. = not significant. D) Equivalent diameter of the hASC 3D cell cultures during on-chip culture. n = 4–12 technical replicates. E) Rates of successful retrieval of the hASC 3D cell cultures from the cell culture chambers. n = 3 biological replicates, * p < 0.05 (one-way ANOVA).

Article Snippet: Human adipose tissue-derived stem cells (hASCs; ASC-F-ZB) were purchased from ZenBio (NC, USA) and maintained in routine 2D cell culture using subcutaneous preadipocyte growth medium (PM-1; ZenBio) according to the supplier's protocol. hASCs at passages 6–8 were used for the experiments.

Techniques: Derivative Assay, Cell Culture

Journal: Cells

Article Title: Short-Term Autophagy Preconditioning Upregulates the Expression of COX2 and PGE2 and Alters the Immune Phenotype of Human Adipose-Derived Stem Cells In Vitro

doi: 10.3390/cells11091376

Figure Lengend Snippet: Autophagy preconditioning in hASCs does not alter self-renewal capacity or surface expression of MSC markers. ( A ) Representative images of hASCs morphology in culture after both short= and long-term exposure to autophagy preconditioning agents. Scale bar = 300 µm. Immunophenotype of Rapa-ASCs ( B ) and 3MA-ASCs ( C ) by flow cytometric analysis of surface proteins. Data are presented as mean ± SEM of 3 independent experiments. Quantifaction and images of colony-forming units by Rapa-ASCs ( D ) and 3MA-ASCs ( E ). Data are presented as mean ± SEM of 4 independent experiments. All data comparing 3 groups are analyzed by using one-way analysis of variance (ANOVA) with Tukey’s post-hoc multiple comparisons, and CFU-F data are analyzed with unpaired student’s t -test. Abbreviations: Rapa, Rapamycin; 3-MA, 3-methyladenine; CFU-F, colony forming units-fibroblasts. ** p < 0.01.

Article Snippet: Primary human ASCs (hASCs) were purchased from LaCell LLC (New Orleans, LA, USA).

Techniques: Expressing

Journal: Cells

Article Title: Short-Term Autophagy Preconditioning Upregulates the Expression of COX2 and PGE2 and Alters the Immune Phenotype of Human Adipose-Derived Stem Cells In Vitro

doi: 10.3390/cells11091376

Figure Lengend Snippet: Autophagy preconditioning in hASCs alters expression of both anti-inflammatory and pro-inflammatory mediators. Rapa-ASCs (left) and 3MA-ASCs (right) were treated for 4 or 24 h, then relative expression levels of anti-inflammatory ( A ) and pro-inflammatory ( B ) genes were measured via RT-qPCR by using the ΔΔCt method. Data are presented as mean relative fold-change ± SEM of 4 independent experiments. ( C ) Secreted PGE2 levels were measured via ELISA in 24 h CM from control and autophagy preconditioned hASCs. Data are presented as means ± SEM of 3 independent experiments. Statistical analysis was performed by using one-way analysis of variance (ANOVA), and differences between the means are indicated with * p < 0.05, ** p < 0.01, *** p < 0.001. Abbreviations: TGF- β , transforming growth factor-beta; IDO , indoleamine 2,3-dioxygenase; TSG-6 , TNF Alpha Induced Protein 6, COX2 , cyclooxygenase 2; IL-6 , interleukin-6; IL-1 β , interleukin-1 beta; PGE2, prostaglandin E2.

Article Snippet: Primary human ASCs (hASCs) were purchased from LaCell LLC (New Orleans, LA, USA).

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control

Journal: Cells

Article Title: Short-Term Autophagy Preconditioning Upregulates the Expression of COX2 and PGE2 and Alters the Immune Phenotype of Human Adipose-Derived Stem Cells In Vitro

doi: 10.3390/cells11091376

Figure Lengend Snippet: Autophagy preconditioning in hASCs alters their transcriptional and secretory response to pro-inflammatory stimulation with hIFNγ. Rapa-ASCs (left) and 3MA-ASCs (right) were preconditioned for 4 or 24 h with their respective autophagy compounds, then stimulated with hIFNγ (5 ng/mL) for 24 h. Following stimulation, anti-inflammatory ( A ) and pro-inflammatory ( B ) genes were measured by using RT-qPCR. Data are presented as means ± SEM of 4 independent experiments and normalized to controls that were not treated with hIFNγ. ( C ) Secreted PGE2 levels were measured in 24 h CM from autophagy-preconditioned and hIFNγ-stimulated hASCs. Data are presented as means ± SEM of 3 independent experiments. All statistical significance is determined by using one-way analysis of variance (ANOVA). Statistical differences between the means are indicated with * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Abbreviations: TGF-β, transforming growth factor-beta; IDO, indoleamine 2,3-dioxygenase; TSG-6, TNF Alpha Induced Protein 6, COX2, cyclooxygenase 2; IL-6, IL-1β, interleukin-6; interleukin-1 beta; PGE2, prostaglandin E2.

Article Snippet: Primary human ASCs (hASCs) were purchased from LaCell LLC (New Orleans, LA, USA).

Techniques: Quantitative RT-PCR

Journal: Regenerative Medicine

Article Title: Therapeutic potential of adipose-derived stem cells and macrophages for ischemic skeletal muscle repair

doi: 10.2217/rme-2016-0094

Figure Lengend Snippet: (A) Mouse bone marrow MPs (mMPs) were co-cultured with mouse ASCs (mASCs) under normoxia (20% oxygen) for 48 h. MPs cultured alone were used as controls (=1). Expression levels of mannose receptor (CD206) were confirmed by flow cytometry. RNA was isolated and gene expression was quantified using SYBR real-time polyamerase chain reaction. (B) mMPs were co-cultured with mASCs under hypoxia (1–2% oxygen) for 48 h. MPs cultured alone were used as controls (=1). Values are displayed as mean ± standard deviation.

Article Snippet: Cell lines Human ASCs (hASCs; PT-5006; Lonza) were commercially purchased and cultured in Dulbecco's Modified Eagle Medium (DMEM; Invitrogen, CA, USA) supplemented with 10% fetal bovine serum (FBS), 1% Glutamax I (Life Tech) and 1% penicillin-streptomycin.

Techniques: Cell Culture, Expressing, Flow Cytometry, Isolation, Gene Expression, Standard Deviation