Journal: Journal of Veterinary Science

Article Title: Toll-like receptor 3-stimulation and aggregate-formation synergistically enhances anti-inflammatory activity of feline mesenchymal stem cells

doi: 10.4142/jvs.23330

Figure Lengend Snippet: IDO-1, indoleamine 2,3-dioxygenase-1; bFGF, basic fibroblast growth factor; TLR3, toll-like receptor 3; fAdMSC, feline adipose-derived mesenchymal stem cell. * p < 0.05.

Article Snippet: TLR3 was stimulated by suspending fAdMSCs were suspended in α-MEM (Nacalai Tesque, Japan) containing 15% fetal bovine serum (HyClone, GE Healthcare UK Ltd., England), penicillin and streptomycin (100 units/mL and 100 μg/mL, respectively, FUJIFILM Wako Pure Chemical Corporation, Japan) (hereafter, culture medium), seeding in wells of a six-well plate (AGC Techno Glass, Japan), and incubating them with a TLR3 ligand (poly [I:C], 1 μg/mL, FUJIFILM Wako Pure Chemical Corporation) for 1 h at 37°C in 5% CO 2 and 95% air.

Techniques: Derivative Assay

Journal: Journal of Veterinary Science

Article Title: Toll-like receptor 3-stimulation and aggregate-formation synergistically enhances anti-inflammatory activity of feline mesenchymal stem cells

doi: 10.4142/jvs.23330

Figure Lengend Snippet: IDO-1, indoleamine 2,3-dioxygenase-1; TLR3, toll-like receptor 3; fAdMSC, feline adipose-derived mesenchymal stem cell.

Article Snippet: TLR3 was stimulated by suspending fAdMSCs were suspended in α-MEM (Nacalai Tesque, Japan) containing 15% fetal bovine serum (HyClone, GE Healthcare UK Ltd., England), penicillin and streptomycin (100 units/mL and 100 μg/mL, respectively, FUJIFILM Wako Pure Chemical Corporation, Japan) (hereafter, culture medium), seeding in wells of a six-well plate (AGC Techno Glass, Japan), and incubating them with a TLR3 ligand (poly [I:C], 1 μg/mL, FUJIFILM Wako Pure Chemical Corporation) for 1 h at 37°C in 5% CO 2 and 95% air.

Techniques: Derivative Assay

Journal: Journal of Veterinary Science

Article Title: Toll-like receptor 3-stimulation and aggregate-formation synergistically enhances anti-inflammatory activity of feline mesenchymal stem cells

doi: 10.4142/jvs.23330

Figure Lengend Snippet: IDO-1, indoleamine 2,3-dioxygenase-1; bFGF, basic fibroblast growth factor; TLR3, toll-like receptor 3; fAdMSC, feline adipose-derived mesenchymal stem cell. * p < 0.05.

Article Snippet: TLR3 was stimulated by suspending fAdMSCs were suspended in α-MEM (Nacalai Tesque, Japan) containing 15% fetal bovine serum (HyClone, GE Healthcare UK Ltd., England), penicillin and streptomycin (100 units/mL and 100 μg/mL, respectively, FUJIFILM Wako Pure Chemical Corporation, Japan) (hereafter, culture medium), seeding in wells of a six-well plate (AGC Techno Glass, Japan), and incubating them with a TLR3 ligand (poly [I:C], 1 μg/mL, FUJIFILM Wako Pure Chemical Corporation) for 1 h at 37°C in 5% CO 2 and 95% air.

Techniques: Derivative Assay

Journal: Journal of Veterinary Science

Article Title: Toll-like receptor 3-stimulation and aggregate-formation synergistically enhances anti-inflammatory activity of feline mesenchymal stem cells

doi: 10.4142/jvs.23330

Figure Lengend Snippet: IDO-1, indoleamine 2,3-dioxygenase-1; bFGF, basic fibroblast growth factor; COX-2, cyclooxygenase-2; TNF-α, tumor necrosis factor-α; LPS, lipopolysaccharide; TLR3, toll-like receptor 3; fAdMSC, feline adipose-derived mesenchymal stem cell. * p < 0.05.

Article Snippet: TLR3 was stimulated by suspending fAdMSCs were suspended in α-MEM (Nacalai Tesque, Japan) containing 15% fetal bovine serum (HyClone, GE Healthcare UK Ltd., England), penicillin and streptomycin (100 units/mL and 100 μg/mL, respectively, FUJIFILM Wako Pure Chemical Corporation, Japan) (hereafter, culture medium), seeding in wells of a six-well plate (AGC Techno Glass, Japan), and incubating them with a TLR3 ligand (poly [I:C], 1 μg/mL, FUJIFILM Wako Pure Chemical Corporation) for 1 h at 37°C in 5% CO 2 and 95% air.

Techniques: Derivative Assay

Journal: Journal of Endotoxin Research

Article Title: Use of murine embryonic fibroblasts to define Toll-like receptor activation and specificity

doi: 10.1177/09680519040100060701

Figure Lengend Snippet: Fig. 1. Murine embryonic fibroblasts (MEFs) express mRNA for TLRs 1–9. MEFs were obtained from wild-type, TLR2 knockout and TLR4 knockout embryos. MEF mRNA was subjected to RT-PCR with primers specific for TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8 and TLR9. PCR products were separated by agarose gel electrophoresis and visualized with ethidium bromide. –RT, control lane without reverse transcriptase. Control lanes (–RT) were negative for all primers; + lane, GAPDH-positive control.

Article Snippet: TLR3 ligand – polyI:C was purchased from Amersham Pharmacia Biotech (Piscataway, NJ, USA) and used at 1 μM concentration.

Techniques: Knock-Out, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Control, Reverse Transcription, Positive Control

Journal: Journal of Endotoxin Research

Article Title: Use of murine embryonic fibroblasts to define Toll-like receptor activation and specificity

doi: 10.1177/09680519040100060701

Figure Lengend Snippet: Fig. 2. TLR2, TLR3 and TLR4 ligands induce cytokine secretion from MEFs. Wild-type MEFs were challenged with LPS (10 ng/ml), Pam3CSK4 (100 ng/ml), Pam2CSK4 (10 ng/ml), IL-1β (100 ng/ml), peptidoglycan (1 µg/ml), zymosan (10 µg/ml) or polyI:C (1 µM) for 18 h. IL-6 and MCP-1 secretion into culture supernatants was analyzed by ELISA. Data are representative of seven cell lines derived from individual embryos and from three cell lines derived from pools of 5–10 embryos.

Article Snippet: TLR3 ligand – polyI:C was purchased from Amersham Pharmacia Biotech (Piscataway, NJ, USA) and used at 1 μM concentration.

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay