human primary liver sinusoid cells  (Axol Bioscience)

Journal: Frontiers in Genetics

Article Title: The role of microRNAs in defining LSECs cellular identity and in regulating F8 gene expression

doi: 10.3389/fgene.2024.1302685

Figure Lengend Snippet: microRNAs targeting F8 gene. (A) Relative expression of F8 gene (Probe ID: ILMN_1675083) in the used ECs and LSECs samples. (B) Left panel: Unsupervised, 3D-PCA displaying different endothelial cells for 25 expressed microRNAs that bind to F8 gene according to IPA. Middle panel: heat map showing the expression of these 25 microRNAs in three LSEC samples. Right panel: Correlation between F8 expression in different endothelial cells and PCA-1, X-axis represents F8 expression and y-axis represents PCA-1. (C) A depiction of the 1808-base pair long 3′ mRNA sequence of the F8 gene along with the 25 expressed microRNAs known to bind to this sequence. (D) Box plots illustrating the expression levels of four microRNAs in different endothelial cells, potentially binding to the F8 gene, as well as the transcription factors binding to the F8 gene promoter. (E) Top Panel: An IPA-generated figure showing the relationships between potentially F8 -binding microRNAs and transcription factors identified using TRANSFAC, which bind to the F8 promoter. Sequence logos of individual transcription factors are displayed on the right. Bottom Panel: Representation of the F8 promoter, covering 1,200 base pairs with 1 kb before the transcription start site and 200 base pairs after the transcription start site. The numbers represent the transcription factors binding sites identified using TRANSFAC (shown and numbered in top panel above).

Article Snippet: In this study, Primary adult Human Liver Sinusoidal Endothelial Cells (LSECs) were procured from Biozol Diagnostica Vertrieb GmbH (product lot No. HEC03020491) and cultured in a specialized endothelial cell culture medium, denoted as ENDO-Growth Medium MED001, supplemented with growth factors and antibiotics.

Techniques: Expressing, Sequencing, Binding Assay, Generated

Journal: Nature Communications

Article Title: Intraocular liver spheroids for non-invasive high-resolution in vivo monitoring of liver cell function

doi: 10.1038/s41467-024-45122-4

Figure Lengend Snippet: a Representative maximum projection of vessels in ACE-liver spheroid acquired by in vivo imaging by intravenous injection of DyLight-649-conjugated lectin (black) after binary conversion and thresholding at 1-week, 1 and 2-months post-tx. Spheroid area limit: pink dashed line. b Immunofluorescence vasculature network (CD31, orange) staining of whole-mount ACE-liver spheroid at 2-months post-tx. Max. projection image. c Vascular density of ACE-liver spheroids. Whiskers represent min/max values, with the mean shown as ‘+’. **** p < 0.0001 by one-way ANOVA test, n = 9 in 3 recipient mice. d Vessel network assessed by number of branch points within the spheroids. Whiskers represent min/max values, with the mean shown as ‘+’. * p < 0.05 by Mann–Whitney test, n = 4 in 3 recipient mice. e Average diameter of iris and intra-spheroid vessels at 1 and 2-months post-tx. Whiskers represent the min/max values, with the mean shown as ‘+’. **** p < 0.0001 by Kruskal–Wallis test, n = 8 measurements per spheroid, 5 spheroids in 3 recipient mice. f In vivo imaging of labeled red blood cells (red) through intra-spheroid vessels (lectin, white) and their trajectory (red-dashed line). Spheroid mass is delimited by a white-dashed line, scale bar = 50 µm. Chronological images taken from Supplementary Movie . g Immunofluorescence staining of whole-mount ACE-liver spheroids and surrounding iris tissue. Sympathetic (TH, red) and parasympathetic (VACHT, green) nerves at 2-months post-tx. Max. projection images. h Immunofluorescence staining within the ACE-liver spheroid mass. Sympathetic (TH, red) and parasympathetic nerves (VACHT, yellow); vessels (CD31, white). Arrowheads indicate nerves alongside vessels and spheroid mass is delimited by white-dashed line. Single plane image. i Immunofluorescence staining of macrophages (F4/80, yellow) within the ACE-liver spheroid. Single plane image. j Experimental outline of human liver spheroids generated from primary human hepatocytes (PHH) and primary human liver sinusoidal endothelial cells (LSECs) transplanted into the ACE of immunodeficient mice. k In vivo imaging of vascularization in engrafted human liver spheroids at 1-month post-tx, visualized by intravenous injection of DyLight-649-conjugated lectin (red). l Vasculature network (CD31, orange) and nuclei (DAPI) staining of ACE-human liver spheroid at 1-month post-tx. Max. projection image. a , b , g , h , i , k , l scale bars = 100 µm. Source data are provided as a Source Data file.

Article Snippet: Cryopreserved purified primary human hepatocytes (PHH) and liver sinusoidal endothelial cells (LSECs) were obtained from suppliers BioIVT (New York, US) and Lonza (Basel, Switzerland), respectively (Supplementary Table ).

Techniques: In Vivo Imaging, Injection, Immunofluorescence, Staining, MANN-WHITNEY, Labeling, Generated