Journal: Stem Cell Research & Therapy

Article Title: Spheroids derived from the stromal vascular fraction of adipose tissue self-organize in complex adipose organoids and secrete leptin

doi: 10.1186/s13287-023-03262-2

Figure Lengend Snippet: AS differentiated with InRo medium have increased expression of adipogenic markers and leptin secretion compared to adipocyte monolayers and increased glycerol secretion in response to isoprenaline stimulation. A Quantification of mRNA levels relative to adult mouse WAT (dotted line). Data are presented as mean ± standard deviation. N = 4 per group. Two-way ANOVA with Tukey's post-test. B Leptin concentration in conditioned medium (48 h) normalized to total DNA. Data are presented as mean ± standard deviation. N = 4 per group. C Differentiated adipocyte monolayers and AS at day 15 of differentiation were incubated with DMEM-F12 medium plus 10% SFB in the absence of insulin and rosiglitazone for 48 h. Next, cells were incubated for additional 48 h with DMEM-F12 medium plus 10% SFB in the presence or absence of 2,000 nM insulin. The conditioned medium (48 h) was harvested for leptin quantification and normalized by total DNA. Data are presented as mean ± standard deviation. N = 4 per group. A significant effect for culture format was determined (F (1, 12) = 21.93; p = 0.0005). D 2D adipocytes and AS were differentiated with InRo protocol and incubated by 3 h in color-free DMEM-F12 medium with or without 1 μM isoprenaline. Conditioned medium was collected for glycerol determination by colorimetric methods and AS. Cells were collected for total DNA extraction and quantification. Data are represented as mean ± standard deviation. N = 3 for monolayer cultures and 5 for AS groups. There was a significant increment in glycerol secretion in response to isoprenaline stimulation (F (1, 11) = 8.357; P = 0.0147). Two-way ANOVA with Tukey's post-test. * p < 0.05; ** p < 0.01; ns: not significant; N.D.: non-detected

Article Snippet: 50–100 AS were incubated for 3 h in color-free DMEM-F12 medium (Gibco, #cat. 21,041,025) with or without 1 μM isoprenaline (Sigma #cat. I5627).

Techniques: Expressing, Standard Deviation, Concentration Assay, Incubation, DNA Extraction

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Patient-derived Mammosphere and Xenograft Tumour Initiation Correlates with Progression to Metastasis

doi: 10.1007/s10911-016-9361-8

Figure Lengend Snippet: Breast cancer mammosphere colony formation and in vivo tumour-initiating activity is increased in metastatic compared to early breast cancers . Cells were isolated from breast cancer samples and grown in suspension culture as mammospheres (representative mammospheres from early (EBC) and metastatic (MBC) samples shown in a ). Metastatic breast cancer samples were more likely to form mammospheres (MS formation >0) than early breast cancers cultured under the same conditions b and had higher primary mammospheres forming efficiencies c (Data presented both as % mammosphere formation, and mammospheres formed/1000 cells plated). No difference in secondary mammosphere formation (self-renewal), defined as a ratio of secondary mammospheres: primary mammospheres, was observed between early and metastatic samples d . Metastatic samples had significantly higher in vivo xenotransplantation potential than early breast cancer samples, over an average period of 200 days, irrespective of tumour phenotype (p = 0.04), where in vivo growth is defined as tumour formed to size limit (1.3cm 3 ) e . Data are presented as mean ± SEM. *p < 0.05 ****p < 0.0005. Statistical analyses: Chi Squared tests (b and e) and two tailed t-test (c and d)

Article Snippet: A single cell suspension was prepared by manual disaggregation (25 gauge needle) and a total of 500 cells/cm 2 were plated in appropriate polyHEMA (Poly (2-hydroxyethylmethacrylate)) coated tissue culture plates in mammosphere medium (phenol red-free DMEM/F12 (Gibco, 21,041)) containing B27 supplement (no vitamin A; Invitrogen, 12,587), rEGF (20 ng/ml; Sigma, E-9644) and Pen-Strep).

Techniques: In Vivo, Activity Assay, Isolation, Cell Culture, Two Tailed Test

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Patient-derived Mammosphere and Xenograft Tumour Initiation Correlates with Progression to Metastasis

doi: 10.1007/s10911-016-9361-8

Figure Lengend Snippet: Mammosphere colony formation predicts metastasis in PDX models. Lung metastases were detected in 14/34 models where a tumour formed in passage 1 by H&E a and confirmed by staining with a human specific mitochondrial antibody b . Lung metastases retained the ER c , PR d and Her2 e status of the primary tumour and contained ki67 positive dividing cells f . Primary mammosphere formation predicted PDX metastases, with samples which metastasised to the lungs in vivo having a significantly higher % mammosphere forming efficiency in vitro g . Metastases are more likely to form in PDX models from high grade EBC tumours (grade 3) than low grade (grade 1 and 2) h . Data are presented as mean ± SEM. * p < 0.05 Statistical analysis: Mann -Whitney U Test

Article Snippet: A single cell suspension was prepared by manual disaggregation (25 gauge needle) and a total of 500 cells/cm 2 were plated in appropriate polyHEMA (Poly (2-hydroxyethylmethacrylate)) coated tissue culture plates in mammosphere medium (phenol red-free DMEM/F12 (Gibco, 21,041)) containing B27 supplement (no vitamin A; Invitrogen, 12,587), rEGF (20 ng/ml; Sigma, E-9644) and Pen-Strep).

Techniques: Staining, In Vivo, In Vitro, MANN-WHITNEY