Project description:The transcriptional program in the response of human fibroblasts to serum. Serum was added to fibroblasts, and samples were taken. This study is described more fully in Iyer VR, et al. 1999. Science 283:83-7 Keywords: time-course
Project description:The transcriptional program in the response of human fibroblasts to serum. Serum was added to fibroblasts in the presence of cycloheximide. This study is described more fully in Iyer VR, et al. 1999. Science 283:83-7 Keywords: time-course
Project description:PI-3K inhibitor (LY294002) was added to quiescent fibroblasts 30 minutes prior growth factor/serum treatments. The role of PI-3K pathway was thus monitored by comparing with untreated samples. Keywords: time course, growth factor response, cell line comparison
Project description:Idiopathic pulmonary fibrosis (IPF) is an incurable lung disease with a poor prognosis. Fibroblasts and myofibroblasts are the key cells in the fibrotic process. Pirfenidone was approved as an anti-fibrotic drug for IPF treatment as it is able to slow disease progression. The mechanisms by which the drug acts on fibroblasts are not clear. Therefore, this study aims to examine the effects of pirfenidone on the human lung fibroblast (HLF) transcriptome in vitro.
Project description:Platelet-rich fibrin (PRF) is prepared from the coagulated plasma of fractionated blood. When squeezing between two plates, PRF is separated into the solid PRF membranes and a liquid exudate, the PRF serum. The question arises regarding the extent to which the overall PRF activity remains in the membranes and what is lost in the serum. To this aim, we have exposed gingival fibroblasts to lysates prepared from PRF membranes and PRF serum, followed by bulk RNA sequencing. A total of 268 up- and 136 down-regulated genes in gingival fibroblasts exposed to PRF lysates are significantly regulated under the premise of a minimum log2 2.5-fold change and a minus log10 significance level of two, respectively. PRF serum caused 62 up- and 32 down-regulated genes when gingival fibroblasts were exposed to PRF serum, respectively. Among the 61 genes commonly up-regulated by PRF lysate and serum were CXCL1, CXCL5, CXCL6, CXCL8, IL33, and IL6 and PTGS2, STC1. PRF lysate further increased the chemokines CCL2, CCL7, CXCL2, CXCL3, and the IL1R1, IL1RL1, and IL1RN – as well as the paracrine factors IL11, LIF, IGF1, BMP2, BMP6, FGF2, CCN2/CTGF and HAS1, HAS2, HAS3. The 16 up-regulated genes by PRF serum included DKK1. Among the 122 down-regulated genes by PRF lysate were IFIT1, IFIT2, IFIT3, OSR1, OSR2. Among the 32 down-regulated genes by PRF serum were FGF18 and GDF15. Taken together, PRF lysates, compared to PRF serum, cause a more complex response of gingival fibroblasts with a chemokine with an obvious increase in chemokine expression and spectrum of paracrine factors.
Project description:We used microarrays to profile gene expression changes following growth factor stimulation of primary human fibroblasts. We serum starved (0.1% serum) fibroblasts for 48 hrs and restimulated with 10% serum for 0, 2, 4, 6 and 8 hrs. Total RNA was extracted from 2 independent biological replicas for each time point and hybridized to expression arrays.
Project description:The transcriptional program in the response of human fibroblasts to serum control group. This study is described more fully in Iyer VR, et al. 1999. Science 283:83-7 Keywords: other
Project description:Normal fibroblasts and SSc fibroblasts between the third and six subpassages were used for experiments. Normal and scleroderma fibroblasts were serum-starved for 24 hours and incubated in the presence or absence of TGF-β1 (2ng/ml) for 6 hours. Total RNA was extracted from culture cells with ISOGEN (Nippon Gene, Tokyo, Japan). MicroRNA isolation from total RNA was performed using RT2 qPCR-Grade miRNA Isolation Kit (SA Bioscience). For RT2 Profiler PCR Array (SABioscience), microRNAs were reverse-transcribed into first strand cDNA using RT2 miRNA First Strand Kit (SABiosciences). A mixture of equal amounts of cDNAs from 5 normal fibroblasts or 5 SSc fibroblasts was prepared. The cDNA was mixed with RT2 SYBR Green/ROX qPCR Master Mix and the mixture was added into a 96-well RT2 miRNA PCR Array (SABiosciences) that included primer pairs for 88 human microRNAs.