Journal: BMC Microbiology

Article Title: First human cell-based cultivation system for the syphilis spirochete Treponema pallidum

doi: 10.1186/s12866-026-04856-5

Figure Lengend Snippet: Human foreskin fibroblast cell lines support the growth of T. pallidum in vitro . A Human foreskin fibroblast cell lines (MoNa, HFF1, and HFFC) support the growth of T. pallidum in long-term, continuous, in vitro cultivation. While cultivation of strain SS14 was discontinued after 10 weeks, in vitro cultures of the DAL-1 strain have been ongoing for over one year. During this period, scheme of subcultures was optimized (up to 1:20). A booster passage (*), extending the standard 7-day subculture to a 14-day period with a cultivation medium exchange on day 7, was employed to enrich the treponemal culture during critical declines. In vitro cultivation was performed in triplicate (i.e., in three cultivation wells) and the mean values for each subculture are presented in graph. Source data from individual wells are shown in Supplementary Table . B Parallel cultivation for seven days ( n = 15) with defined DAL-1 inoculum (10 6 treponemes) validated the differences in growth support of individual foreskin cell lines. Treponemal growth on human foreskin fibroblast cells was slower compared to growth on rabbit epithelial Sf1Ep cells. Red bar, mean. C Human foreskin-based cultivation system supports the growth of various T. pallidum strains ( n = 8), from the Nichols-like as well as the SS14-like cluster. Note that human foreskin fibroblast cells were prepared as an equal mixture of three tested cell lines. Each T. pallidum strain was cultivated in a single in vitro well, representing a sole experimental replicate used for data acquisition

Article Snippet: Human foreskin fibroblasts HFF1 (SCRC-1041; ATCC) and HFFC (300715; Cytion) were purchased, while the third cell line (MoNa) was kindly provided by Dr. Vladimir Rotrekl (Masaryk University), and was originally obtained from the National Tissue Centre (Czech Republic).

Techniques: In Vitro

Journal: Pharmaceuticals

Article Title: Ethanolic Fenugreek Extract: Its Molecular Mechanisms against Skin Aging and the Enhanced Functions by Nanoencapsulation

doi: 10.3390/ph15020254

Figure Lengend Snippet: Anti-aging activities’ evaluation of fenugreek extract: ( a ) In vitro collagenase inhibition of fenugreek extract (denoted as Extract) and liponiosome encapsulating fenugreek extract (denoted as LNF). ( b ) Effect of fenugreek extract on collagen production. Human dermal fibroblast cells were treated with 125 µg/mL of fenugreek extract and rutin, and 50 µg/mL of vitamin C as a positive control and 0.005% DMSO as a control (vehicle), for 7 and 14 days. Data are reported as means ± SD ( n = 3). *, p < 0.05; ***, p < 0.001.

Article Snippet: Human dermal fibroblast cells were purchased from the American Type Culture Collection: ATCC (PCS-201-010) and immortalized human keratinocytes (HaCaT) were purchased from Cell Lines Service, Germany (Cat. No. 300493).

Techniques: In Vitro, Inhibition, Positive Control, Control

Journal: Pharmaceuticals

Article Title: Ethanolic Fenugreek Extract: Its Molecular Mechanisms against Skin Aging and the Enhanced Functions by Nanoencapsulation

doi: 10.3390/ph15020254

Figure Lengend Snippet: Evaluation of LNF activities: ( a ) Cytotoxicity induced by LNF: The effect of blank, LNF, and fenugreek extract on cell viability. Human dermal fibroblast cells were treated with different concentrations of blank (liponiosome without fenugreek extract denoted as Blank), LNF, and fenugreek extract for 24 h. Data are means ± SD ( n = 3). ( b ) Collagen production induced by LNF: The effect of LNF on collagen production. Human dermal fibroblast cells were treated with 0.005% DMSO as a control (vehicle), 7 µg/mL of extract, and 100 µg/mL of LNF (7 µg/mL of extract equivalence), together with blank particles, for 7 and 14 days. Data are represented as means ± SD ( n = 3). *, p < 0.05.

Article Snippet: Human dermal fibroblast cells were purchased from the American Type Culture Collection: ATCC (PCS-201-010) and immortalized human keratinocytes (HaCaT) were purchased from Cell Lines Service, Germany (Cat. No. 300493).

Techniques: Control

Journal: Pharmaceuticals

Article Title: Ethanolic Fenugreek Extract: Its Molecular Mechanisms against Skin Aging and the Enhanced Functions by Nanoencapsulation

doi: 10.3390/ph15020254

Figure Lengend Snippet: Inhibition of UV-induced MMPs and interleukin secretion on co-cultured skin cells by fenugreek extract and LNF: ( a ) Effect of UV-induced cytotoxicity after the pretreatments of resveratrol, rutin, fenugreek extract, blank nanoparticles (liponiosome without fenugreek extract), LNF nanoparticles, and 0.005% DMSO as a control (vehicle). HaCAT and human dermal fibroblast cells were co-cultured and pretreated with 7 µg/mL of extract and 100 µg/mL of LNF (7 µg/mL of extract equivalence), together with 100 µg/mL of blank, 10 µg/mL of resveratrol, 7 µg/mL of rutin as a positive, and 0.005% of DMSO as a control (vehicle), for 24 h before UV exposure. Data are means ± SD ( n = 3). *, p < 0.05. ( b ) The levels of UV-induced MMP1 and MMP9 secretions after fenugreek extract and LNF treatments. Data are reported as means ± SD ( n = 3). *, p < 0.05. ( c ) The levels of UV-induced IL-6 and IL-8 secretions after fenugreek extract and LNF treatments. Data are reported as means ± SD ( n = 3). *, p < 0.05.

Article Snippet: Human dermal fibroblast cells were purchased from the American Type Culture Collection: ATCC (PCS-201-010) and immortalized human keratinocytes (HaCaT) were purchased from Cell Lines Service, Germany (Cat. No. 300493).

Techniques: Inhibition, Cell Culture, Control

Journal: SLAS discovery : advancing life sciences R & D

Article Title: Patient-Derived Phenotypic High-Throughput Assay to Identify Small Molecules Restoring Lysosomal Function in Tay–Sachs Disease

doi: 10.1177/2472555218814538

Figure Lengend Snippet: Development of a lysosomal calcium release assay in TSD patient-derived fibroblasts. Representative kinetic tracings from WT (A) and TSD (B) fibroblast calcium release in response to GPN measured on the FDSS μCell. Cells were pretreated with 25 μM APB for 15 min prior to GPN addition. Tracings show the baseline recording starting the last 5 min of the APB pretreatment. Ionomycin (5 μM) was added to induce a maximal release from the ER for comparison once the GPN-induced calcium release reached baseline. (C) Peak fluorescence ratios measured across a range of GPN concentrations (6–100 μM) in WT and TSD fibroblasts. The S/B indicated for each GPN concentration was determined comparing the peak fluorescence ratio calcium response in WT (signal) with TSD (background) fibroblasts. A 50 μM concentration of GPN was selected for the screening assay protocol as it produced the best S/B. (D) GPN-induced calcium release from acidic lysosomal calcium stores is significantly reduced in fibroblasts derived from TSD patients. Patient cell lines are WT (GM05659), TSD 1 (GM00221), TSD 2 (GM00502), and TSD 3 (GM11853). ***p < 0.0001 compared with WT cells. The reduced lysosomal calcium response in TSD patient cells was used as the basis to develop the phenotypic HTS assay.

Article Snippet: Human fibroblast cell lines from Tay–Sachs patients (GM00221, GM00502, GM11853) and a healthy normal patient (GM05659) were from Coriell Institute for Medical Research (Camden, NJ); GM00221 donor is homozygous for a 4-base-pair insertion at nucleotide 1278 in exon 11 of the HEXA gene (c.1278insTATC), which leads to a premature termination signal.

Techniques: Release Assay, Derivative Assay, Comparison, Fluorescence, Concentration Assay, Screening Assay, Produced, HTS Assay

Journal: SLAS discovery : advancing life sciences R & D

Article Title: Patient-Derived Phenotypic High-Throughput Assay to Identify Small Molecules Restoring Lysosomal Function in Tay–Sachs Disease

doi: 10.1177/2472555218814538

Figure Lengend Snippet: Autophagy detection in WT and TSD fibroblasts. (A) Immunoblot analyses with anti-LC3 and anti-actin antibodies on WT (GM05659) and TSD (GM00221) patient fibroblast cell lines. Cells were treated with either vehicle (0.2% DMSO), 20 μM pyrimethamine (PYR), 100 nM bafilomycin A1 (Baf. A1), or PYR plus Baf. A1 for 22 h, followed by Western blot analysis. A representative Western blot is shown. (B) Quantification of LC3II intensity normalized to β-actin. Data represent the mean + SEM of at least three independent replicates.

Article Snippet: Human fibroblast cell lines from Tay–Sachs patients (GM00221, GM00502, GM11853) and a healthy normal patient (GM05659) were from Coriell Institute for Medical Research (Camden, NJ); GM00221 donor is homozygous for a 4-base-pair insertion at nucleotide 1278 in exon 11 of the HEXA gene (c.1278insTATC), which leads to a premature termination signal.

Techniques: Western Blot