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: Immunopharmacology and immunotoxicology

Article Title: Circular RNA circCTNNA1 is downregulated in osteoarthritis and sponges miR-29a to suppress LPS-induced apoptosis of synoviocytes.

doi: 10.1080/08923973.2021.1988103

Figure Lengend Snippet: Figure 2. CircCTNNA1 and miR-29a directly interacted with each other in synoviocytes. The interaction between circCTNNA1 and miR-29a was predicted by IntaRNA2.0 (A) and analyzed using RNA pull-down assay (B). The interaction between circCTNNA1 (mut) and miR-29a was also analyzed by RNA pull-down assay (C). p< .05.

Article Snippet: Synoviocytes (type B, primary cells) derived from an adult OA patient were purchased from Sigma-Aldrich (Cat. 408OA-05A) and cultured in human synoviocyte media (Cell applications) at 37 C in an incubator with 5% CO2 and 95% humidity.

Techniques: Pull Down Assay

Journal: Immunopharmacology and immunotoxicology

Article Title: Circular RNA circCTNNA1 is downregulated in osteoarthritis and sponges miR-29a to suppress LPS-induced apoptosis of synoviocytes.

doi: 10.1080/08923973.2021.1988103

Figure Lengend Snippet: Figure 3. CircCTNNA1 and miR-29a failed to regulate the expression of each other in synoviocytes. To explore the interaction between circCTNNA1 and miR-29a, synoviocytes were transfected with either circCTNNA1 expression vector or miR-29a mimic and the overexpression was confirmed every 24 h until 96 h (B). The effects of circCTNNA1 and miR-29a overexpression on the expression of each other were analyzed by RT-qPCR (B). The correlations between circCTNNA1 and miR- 29a were analyzed by Pearson’s correlation coefficient across both OA (C) and control (D) synovial fluid samples. p < .05.

Article Snippet: Synoviocytes (type B, primary cells) derived from an adult OA patient were purchased from Sigma-Aldrich (Cat. 408OA-05A) and cultured in human synoviocyte media (Cell applications) at 37 C in an incubator with 5% CO2 and 95% humidity.

Techniques: Expressing, Transfection, Plasmid Preparation, Over Expression, Quantitative RT-PCR, Control

Journal: Immunopharmacology and immunotoxicology

Article Title: Circular RNA circCTNNA1 is downregulated in osteoarthritis and sponges miR-29a to suppress LPS-induced apoptosis of synoviocytes.

doi: 10.1080/08923973.2021.1988103

Figure Lengend Snippet: Figure 4. CircCTNNA1 may sponge miR-29a to reduce the apoptosis of synoviocytes induced by LPS. Synoviocytes were treated with LPS at dosages of 0, 2, 4, 6, 8, and 10 lg/ml for 48 h, and the expression of circCTNNA1 (A) and miR-29a (B) was determined by RT-qPCR. The role of circCTNNA1 and miR-29a in regulating the apoptosis of synoviocytes induced by LPS (10 lg/ml for 48 h) was analyzed by cell apoptosis (C). Expression of active (cleaved) caspase-3 in different groups was analyzed by Western blot (D). The role of circCTNNA1 (mut) in cell apoptosis was also analyzed (E). p < .05.

Article Snippet: Synoviocytes (type B, primary cells) derived from an adult OA patient were purchased from Sigma-Aldrich (Cat. 408OA-05A) and cultured in human synoviocyte media (Cell applications) at 37 C in an incubator with 5% CO2 and 95% humidity.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

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