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Human hepatic stellate cells

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Human hepatic stellate cells are a type of mesenchymal cells found in the liver. They play a key role in the maintenance and repair of the liver's extracellular matrix.

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Lab products found in correlation

Opera phenix high content imaging platform, by PerkinElmer (1 mentions) Anti αsma clone 1a4, by Merck Group (1 mentions) Anti col1a1, by Merck Group (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) Tgf β1, by R&D Systems (1 mentions) 1640 medium, by Merck Group (1 mentions) Fetal calf serum (fcs), by Merck Group (1 mentions) Dm1000, by Leica (1 mentions) Hibec, by ScienCell (1 mentions) Tgf β, by Merck Group (1 mentions) Mg132, by Cell Signaling Technology (1 mentions) Bafilomycin a1, by Merck Group (1 mentions) Lipofectamine 3000 kit, by Thermo Fisher Scientific (1 mentions) Recombinant tgf β, by R&D Systems (1 mentions)

4 protocols using human hepatic stellate cells

1

Inhibition of Hepatic Stellate Cell Fibrogenesis

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The effect of PF-05221304 to inhibit activation of hepatic stellate cells to a fibrogenic myofibroblast was assessed by measuring the effect of the compound on the production of αSMA and collagen from primary hepatic stellate cells stimulated with TGFβ1 to drive fibrogenesis. Briefly, human hepatic stellate cells (cat 5300; Sciencell Research Laboratories, Carlsbad, CA) were treated with 1 ng/mL TGFβ1 (cat 240B; R&D Systems, Minneapolis, MN) in the presence of varying concentrations of PF-05221304 or vehicle. Ascorbic acid was added at 100 μmol/L to the cell culture media to enhance the collagen production. Hepatic stellate cells unstimulated with TGFβ1 were used as a control. After a 72-hour incubation, the wells were methanol fixed, blocked, and incubated with anti-αSMA (clone 1A4; Sigma Aldrich) and anti-Col1A1 (cat C2456; Sigma), followed by matching secondary antibodies and Hoechst (Invitrogen, Carlsbad, CA) to produce an immunofluorescence signal. The cells were imaged on an Opera Phenix High Content Imaging platform (PerkinElmer). Images were analyzed with custom scripts in Columbus (PerkinElmer) for αSMA and collagen fluorescence intensity and nuclear counts. Effects on TGFβ1-stimulated proliferation was assessed by comparing the nuclear cell density of compound-treated cells to TGFβ1-stimulated vehicle-treated cells and the unstimulated control cells.
Ross T.T., Crowley C., Kelly K.L., Rinaldi A., Beebe D.A., Lech M.P., Martinez R.V., Carvajal-Gonzalez S., Boucher M., Hirenallur-Shanthappa D., Morin J., Opsahl A.C., Vargas S.R., Bence K.K., Pfefferkorn J.A, & Esler W.P. (2020). Acetyl-CoA Carboxylase Inhibition Improves Multiple Dimensions of NASH Pathogenesis in Model Systems. Cellular and Molecular Gastroenterology and Hepatology, 10(4), 829-851.
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2

Immortalized Cell Lines for Liver Research

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The immortalized normal human cholangiocyte cell line, H69, was a gift from Dr. G.J. Gores, Mayo Clinic, Rochester, MN. Immortalized murine normal pooled cholangiocyte lines were also used.(10 (link)) Human LSCs were purchased from Creative Bioarray (Shirley, NY). The human hepatic stellate cells,(20 (link)) human mesenchymal stem cells (MSCs) and human hepatocytes (HHs) were purchased from Sciencell (Carlsbad, CA). The human acute monocytic leukemia cell line (THP-1) was purchased from ATCC Inc. (Manassas, VA) and maintained in Roswell Park Memorial Institute 1640 medium (Sigma-Aldrich) with 10% fetal calf serum (FCS) without antibiotics.
McDaniel K., Wu N., Zhou T., Huang L., Sato K., Venter J., Ceci L., Chen D., Ramos-Lorenzo S., Invernizzi P., Bernuzzi F., Wu C., Francis H., Glaser S., Alpini G, & Meng F. (2019). Amelioration of Ductular Reaction by Stem Cell Derived Extracellular Vesicles in MDR2 Knockout Mice via Lethal-7 microRNA. Hepatology (Baltimore, Md.), 69(6), 2562-2578.
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3

Wound Healing Assay with HSCs

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Human hepatic stellate cells (#5300; ScienCell, Carlsbad, CA) were plated in a poly-L-lysine-coated 24-well plate and cultured until confluence. A gap was created by scratching the cell layer, and the stimulation was initiated. The area of the scratch was measured repeatedly over time via bright-field microscopy (DM1000; Leica Microsystems, Wetzlar, Germany).
Reißing J., Berres M., Strnad P., Wree A., Inzaugarat M.E., Trautwein C., Bruns T, & Zimmermann H.W. (2023). Th2 Cell Activation in Chronic Liver Disease Is Driven by Local IL33 and Contributes to IL13-Dependent Fibrogenesis. Cellular and Molecular Gastroenterology and Hepatology, 17(4), 517-538.
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4

Investigating TGFβ-Mediated Signaling in Cholangiocyte and Hepatic Cells

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The H69 cholangiocyte cell line was provided by Nicholas LaRusso, M.D. (Mayo Clinic). Human Intrahepatic Biliary Epithelial Cells (HiBEC) and Human Hepatic Stellate Cells (hHSC) were purchased from ScienCell (Catalog #5100 and #5300, respectively). The RGF-1 portal fibroblast cell line was provided by Jonathan Dranoff, M.D. (University of Arkansas) (17 ). Cells were grown in DMEM/F12 supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin, adenine, insulin, epinephrine, T3-T, hydrocortisone, and epidermal growth factor. Cells were serum starved in basal DMEM containing 1% penicillin/streptomycin for 4 hours or overnight before treatment with 10 ng/mL recombinant TGFβ (R&D Systems #240-B). In some experiments, cells were incubated with 10µM MG132 (Cell Signaling Technology #2194), 10 µM E64D (Sigma Aldrich #E8440), or 10 µM Bafilomycin A1 (Sigma-Aldrich #B1793), one hour prior to TGFβ. For some experiments, cells at 70% confluence were transfected using the Lipofectamine 3000 kit (Invitrogen #L3000–015). The plasmids used for transfection were from Addgene; HA-Ubiquitin (WT, #17608, K48, # 17605, K63, #17606), myc-EZH2 WT was obtained from William Faubion, M.D. (Mayo Clinic).
Jalan-Sakrikar N., De Assuncao T.M., Shi G., Aseem S., Chi C., Shah V.H, & Huebert R.C. (2019). Proteasomal Degradation of Enhancer of Zeste Homologue 2 in Cholangiocytes Promotes Biliary Fibrosis. Hepatology (Baltimore, Md.), 70(5), 1674-1689.
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