Project description:Gene expression profiling of HUVEC (human umbilical vein EC cell; Lonza), HAEC (human aortic EC cells), HCAEC (human coronary artery EC cells), HPAEC (human pulmonary artery EC cells), HMVEC (human microvascular (dermal) , HASMC ( Human Aortic Smooth Muscle Cells), T cells and Bcells. Gene expression profiling of Endothelial cells and Non-endothelial cells in order to identify the genes with preferntial expression to endothelial cells. The experiments are performed in duplicate on both the HT Human Genome U133A and U133B arrays.
Project description:DNA microarrays were used to investigate global gene expression patterns in cultured human umbilical artery endothelial cells (HUAECs) exposed to 1 nmol/L estradiol and/or 100 µg/ml oxidized low density lipoprotein (oxLDL) for 24 hours compared to control cells.
Project description:To study the impact of the organotypic assembly of vascular smooth muscle cells on their transcriptome, we cultured human umbilical artery smooth muscle cells under 2D conditions and as aggregates in hanging drops under 3D conditions. After 48 hours, RNA was isolated from both groups
Project description:Umbilical vein, lung microvascular, aortic and coronary artery endothelial cell profiles generated. Further characterization gained by comparison with other selected cell types. Keywords: other
Project description:Dysfunctional umbilical cord blood (CB) is an important factor for the development of IUGR in utero. However, the genetic mechanism underlying the miRNAs in CB exosomes influence the development of IUGR is not well characterized. Herein, we present a comprehensive investigation of miRNA transcriptome of umbilical cord vein and artery between IUGR and normal littermate. We total identified 636 unique miRNAs. There were 116 significant differentially expressed (DE) miRNAs between umbilical vein of normal (NV) and IUGR (IV) and 226 DE miRNAs between umbilical artery of normal (NA) and IUGR (IA) (P < 0.001). Cluster analysis revealed that umbilical artery had the most striking divergence, implied it plays more prominent role in the development of IUGR. The miRNAs enriched in NA mainly participate in blood vessel development, regulation of transcription and growth. The miRNAs highly expressed in IA mainly enriched in apoptosis, cell death, embryonic development and immune system development Besides, miRNAs related to oxygen transfer (miR-210, miR-424), angiogenesis (miR-130a, miR-150, miR-34a) and immune system development (miR-181a, miR-155) were lower expressed in IUGR. Our findings demonstrate that CB derived miRNAs participate in fetal epigenetic regulation during pregnancy, which may supply a new explanation for abnormal embryologic development and some congenital diseases.
Project description:Endothelial cells (EC) lining arteries and veins have distinct molecular and functional signatures. The (epi)genetic regulatory mechanisms underlying this heterogeneity in human EC are incompletely understood. Using genome-wide microarray screening we established a specific fingerprint of freshly isolated arterial (HUAEC) and venous EC (HUVEC) from human umbilical cord comprising 64 arterial and 12 venous genes, representing distinct functions and pathways. Among the arterial genes were 8 transcription factors, including HEY2, a downstream target of Notch signaling and the current ‘golden standard’ pathway for arterial EC specification. Short-term culture of HUAEC or HUVEC abrogated differential gene expression resulting in a default state. Erasure of arterial gene expression was at least in part due to loss of canonical Notch activity and HEY2 expression. Notably, nCounter analysis revealed that restoring HEY2 expression or Delta-like 4 (Dll4)-induced Notch signaling in cultured HUVEC or HUAEC only partially reinstated the arterial EC gene signature while combined overexpression of the 8 transcription factors restored this fingerprint much more robustly. Each transcription factor had a different impact on gene regulation, with some stimulating only few and others boosting a large proportion of arterial genes. Interestingly, although there was some overlap and cross-regulation, the transcription factors largely complemented each other in regulating the arterial EC gene profile. Thus, our study showed that Notch signaling determines only part of the arterial EC signature and identified additional novel and complementary transcriptional players in the complex regulation of human arteriovenous EC identity To identify an arteriovenous (AV) fingerprint in human endothelial cells (EC) across different vascular beds, we used microarrays on RNA from 38 EC samples corresponding to 6 cultured human arterial-EC types (hepatic artery EC or HHAEC, N=3; aorta EC or HAEC, N=2; coronary artery EC or HCAEC, N=2; iliac artery EC or HIAEC, N=2; pulmonary artery EC or HPAEC, N=3; and umbilical artery EC or HUAEC-C, N=5), 4 cultured human venous-EC types (hepatic vein EC or HHVEC, N=3; iliac vein EC or HIVEC, N=3; pulmonary vein EC or HPVEC, N=2; and umbilical vein EC or HUVEC-C, N=5), freshly isolated HUAEC (HUAEC-F, N=4) and freshly isolated HUVEC (HUVEC-F, N=4). Due to the difficulty to obtain biopsies from healthy donors, we did not have access to freshly isolated aEC or vEC matched for all cultured EC types.
Project description:Umbilical vein, lung microvascular, aortic and coronary artery endothelial cell profiles generated. Further characterization gained by comparison with other selected cell types.
Project description:To study the impact of TGF signaling on the transcriptome of vascular smooth muscle cells, we cultured human umbilical artery smooth muscle cells as 3D aggregates in hanging drops for 48 hours. One group was treated with TGFβRI/II inhibitor LY2109761 (10 μM, additional 10 μM after 24 h), the other group with solvent (DMSO) as control. RNA was isolated from both groups after 48h