Molecular Analysis of Fetal and Adult Primary Human Liver Sinusoidal Endothelial Cells: A Comparison to Other Endothelial Cells

Abstract

In humans, Factor VIII (F8) deficiency leads to hemophilia A and F8 is largely synthesized and secreted by the liver sinusoidal endothelial cells (LSECs). However, the specificity and characteristics of these cells in comparison to other endothelial cells is not well known. In this study, we performed genome wide expression and CpG methylation profiling of fetal and adult human primary LSECs together with other fetal primary endothelial cells from lung (micro-vascular and arterial), and heart (micro-vascular). Our results reveal expression and methylation markers distinguishing LSECs at both fetal and adult stages. Differential gene expression of fetal LSECs in comparison to other fetal endothelial cells pointed to several differentially regulated pathways and biofunctions in fetal LSECs. We used targeted bisulfite resequencing to confirm selected top differentially methylated regions. We further designed an assay where we used the selected methylation markers to test the degree of similarity of in-house iPS generated vascular endothelial cells to primary LSECs; a higher similarity was found to fetal than to adult LSECs. In this study, we provide a detailed molecular profile of LSECs and a guide to testing the effectiveness of production of in vitro differentiated LSECs.

Cell-free, methylated DNA in blood samples reveals tissue-specific, cellular damage from radiation treatment

“…We thus generated tissue-specific endothelial methylomes paired with transcriptomes to be able to identify damage to distinct populations of microvascular and large vessel endothelial cell types including coronary artery, pulmonary artery, cardiac microvascular, and liver sinusoidal endothelia. We also made use of publicly available liver sinusoidal endothelial(52) and umbilical vein endothelial methylomes(53) to complement our data ( Supplemental Table S1 ). Previous studies support considering the heart and lung as an integrated system in the development of radiation damage due to the shared cardiopulmonary circulation(4).…”

Membrane Properties of Human Induced Pluripotent Stem Cell-Derived Cultured Red Blood Cells

“…Five human iPSC lines derived from erythroblasts or CB CD34+ cells were used [ 9 , 13 , 14 , 15 ] ( Supplementary Table S1 ). Native RBCs were obtained from RBC units within 24 h after donation, and nRETs were isolated from CB with magnetic beads within 12 h postpartum (CD71 Microbead Kit; Miltenyi Biotec, Bergisch Gladbach, Germany).…”

A Review of the Evidence for Tryptophan and the Kynurenine Pathway as a Regulator of Stem Cell Niches in Health and Disease

“… 138 Until recently most studies investigating KP expression using primary human cells have been limited to those using easy-to-access, minimally-invasive sources of cells, like blood 3 or using developing brain cells, 4 , 6 - 10 , 63 , 139 however potential differences in signaling system expression upon differentiation and maturation of cells, and also between organism embryonic or foetal development (stem/precursor cells within), and adult stem cells residing in niches and iPS-derived cells, need to be appreciated and not assumed there is no difference. The use of high-throughput transcriptomics and other genetic approaches has helped researchers in various fields towards answering these questions, 140 - 142 however it should be recognised that there can be intrinsic biases favouring detection of highly expressed transcripts at the expense of modestly-expressed ones; and basal KP expression in many cells may fall into this latter category. Also, RNA from heterogenously-expressing samples/derived from multiple cell types will essentially be averaged in output datasets.…”