Journal: Microbial cell factories

Article Title: Secretion of tumoricidal human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) by recombinant Lactococcus lactis: optimization of in vitro synthesis conditions.

doi: 10.1186/s12934-018-1028-2

Figure Lengend Snippet: Fig. 9 Selectivity of hsTRAIL-induced death to cancer cells. HCT116 cells and human cardiac fibroblasts were incubated for 48 h with increasing concentration of hsTRAIL present in supernatant from broth culture of L. lactis (hsTRAIL+). As controls in experimental setup were used: corresponding volumes of supernatants from broth culture of L. lactis (hsTRAIL−)—negative control; corresponding concentrations of recombinant human TRAIL (Peprotech)—positive control. Viability of cancer cells and non-malignant, cardiac fibroblasts was assessed by MTS assay. Results are presented as % of viability of cells incubated in standard culture medium only. Secreted hsTRAIL remained non-cytotoxic to fibroblasts (Fig. 9a), while decreased viability of cancer cells in a dose-dependent manner (Fig. 9b). Table below—concentration of hsTRAIL [ng/ml] in specified volume of supernatant. The bars indicate the mean value ± SD of three independent experiments, each performed in triplicates. Statistical significance was calculated using two-way ANOVA with Tukey’s multiple comparison post-test. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Human primary proliferating cardiac fibroblasts were obtained from Cell Applications, Inc. (San Diego, CA).

Techniques: Incubation, Concentration Assay, Negative Control, Recombinant, Positive Control, MTS Assay, Comparison

Journal: GeroScience

Article Title: Rapamycin increases oxidative metabolism and enhances metabolic flexibility in human cardiac fibroblasts

doi: 10.1007/s11357-018-0030-2

Figure Lengend Snippet: Progerin expression induces a glycolytic phenotype which is reversed by rapamycin. Measurements of basal respiration (a), glucose uptake (b), and extracellular acidification (ECAR) (c) in human cardiac fibroblasts (HCFs) expressing lamin A or progerin maintained with or without 1 nM rapamycin. Oxygen consumption rates were determined using a Seahorse bioanalyzer. Oxygen consumption and extracellular acidification rates were normalized to cell number. Data presented as mean ± SD (n = 2–3, in triplicates). The differences are shown to be statistically significant with p < 0.05 in comparison to the late passage + rapamycin values

Article Snippet: Cell culture and cell culture reagents Human cardiac fibroblast cells (ScienCell Inc. Carlsbad, CA) and HGPS patient-derived fibroblasts (Coriell Institute, Camden, NJ) were cultivated in minimal essential medium (MEM) supplemented with 10% fetal bovine serum, 1% l -glutamine, 1% MEM vitamins, and 1% MEM non-essential amino acids according to standard culture protocol for lifespan analysis of human diploid fibroblasts (Cristofalo and Charpentier 1980 ), with or without the addition of 1 nM rapamycin.

Techniques: Expressing, Comparison

Journal: GeroScience

Article Title: Rapamycin increases oxidative metabolism and enhances metabolic flexibility in human cardiac fibroblasts

doi: 10.1007/s11357-018-0030-2

Figure Lengend Snippet: Late passage human fibroblasts are relatively glycolytic. Seahorse bioanalyzer measurements of basal respiration (a), glucose uptake (b), and extracellular acidification (c) in early and late passage (> 90% lifespan completed) human cardiac fibroblasts. The values were normalized to cell numbers in each well. Samples were measured in quintuplicate and results are representative of at least two independent experiments ± SD. The bars with an asterisk represent values that are significantly (*, p < 0.05) differ from the late passage ones

Article Snippet: Cell culture and cell culture reagents Human cardiac fibroblast cells (ScienCell Inc. Carlsbad, CA) and HGPS patient-derived fibroblasts (Coriell Institute, Camden, NJ) were cultivated in minimal essential medium (MEM) supplemented with 10% fetal bovine serum, 1% l -glutamine, 1% MEM vitamins, and 1% MEM non-essential amino acids according to standard culture protocol for lifespan analysis of human diploid fibroblasts (Cristofalo and Charpentier 1980 ), with or without the addition of 1 nM rapamycin.

Techniques:

Journal: GeroScience

Article Title: Rapamycin increases oxidative metabolism and enhances metabolic flexibility in human cardiac fibroblasts

doi: 10.1007/s11357-018-0030-2

Figure Lengend Snippet: Distinct rates of pyruvate and glutamine/palmitate oxidation by early and late passage human fibroblasts and the effect of rapamycin. a The rates of pyruvate-mediated respiration were calculated as the percentage of inhibition of oxygen consumption by specific pyruvate transporter inhibitor UK5099. The rates of oxygen consumption obtained via sequential addition of first the glutamine and palmitate oxidation inhibitors, BPTES and etomoxir, and then UK5099 represent the capacity of pyruvate oxidation. b The rates of glutamine/palmitate oxidation were determined in the presence of corresponding inhibitors BPTES and etomoxir, while the capacity was determined following first the addition of pyruvate inhibitor UK5099 and then BPTES/etomoxir. c, d Effect of rapamycin on capacity of late passage cells to oxidize pyruvate and glutamine/palmitate in late passage fibroblasts maintained in the presence or absence of 1 nM rapamycin. Values are generated as in b through the addition of inhibitors and calculating the percent inhibition of oxygen consumption. In all cases, graphs represent the values of one of at least two independent experiments measured in triplicates and presented as mean ± SD; *p < 0.05 vs late passage

Article Snippet: Cell culture and cell culture reagents Human cardiac fibroblast cells (ScienCell Inc. Carlsbad, CA) and HGPS patient-derived fibroblasts (Coriell Institute, Camden, NJ) were cultivated in minimal essential medium (MEM) supplemented with 10% fetal bovine serum, 1% l -glutamine, 1% MEM vitamins, and 1% MEM non-essential amino acids according to standard culture protocol for lifespan analysis of human diploid fibroblasts (Cristofalo and Charpentier 1980 ), with or without the addition of 1 nM rapamycin.

Techniques: Inhibition, Generated

Journal: GeroScience

Article Title: Rapamycin increases oxidative metabolism and enhances metabolic flexibility in human cardiac fibroblasts

doi: 10.1007/s11357-018-0030-2

Figure Lengend Snippet: Rapamycin improves metabolic flexibility in human fibroblast cells expressing progerin. Measurements of the capacity for oxidizing pyruvate (a) or glutamine/palmitate (b) in late passage fibroblasts maintained in the presence or absence of 1 nM rapamycin. Measurements were made as in Fig. ​Fig.2.2. The data were normalized to cell number in each well. Data presented as mean ± SD (n = 2, in triplicates). The differences are shown to be statistically significant with p < 0.05 in comparison to the late passage values

Article Snippet: Cell culture and cell culture reagents Human cardiac fibroblast cells (ScienCell Inc. Carlsbad, CA) and HGPS patient-derived fibroblasts (Coriell Institute, Camden, NJ) were cultivated in minimal essential medium (MEM) supplemented with 10% fetal bovine serum, 1% l -glutamine, 1% MEM vitamins, and 1% MEM non-essential amino acids according to standard culture protocol for lifespan analysis of human diploid fibroblasts (Cristofalo and Charpentier 1980 ), with or without the addition of 1 nM rapamycin.

Techniques: Expressing, Comparison

Journal: GeroScience

Article Title: Rapamycin increases oxidative metabolism and enhances metabolic flexibility in human cardiac fibroblasts

doi: 10.1007/s11357-018-0030-2

Figure Lengend Snippet: Rapamycin prevents premature senescence induced by galactose. Human cardiac fibroblasts maintained with or without 1 nM rapamycin were transferred to glucose-free medium supplemented with either 10 or 20 mM galactose at a population doubling of 27.5. Cumulative population doublings were calculated as described in “Materials and methods” section

Article Snippet: Cell culture and cell culture reagents Human cardiac fibroblast cells (ScienCell Inc. Carlsbad, CA) and HGPS patient-derived fibroblasts (Coriell Institute, Camden, NJ) were cultivated in minimal essential medium (MEM) supplemented with 10% fetal bovine serum, 1% l -glutamine, 1% MEM vitamins, and 1% MEM non-essential amino acids according to standard culture protocol for lifespan analysis of human diploid fibroblasts (Cristofalo and Charpentier 1980 ), with or without the addition of 1 nM rapamycin.

Techniques: