The supernatant was centrifuged at 600 × g to obtain hepatic nonparenchymal cells. Primary HSCs were isolated by density-gradient centrifugation using ficoll paque-plus (GE Healthcare, Madison, WI) and percoll (GE Healthcare, Madison, WI, USA)41 (link). Non-parenchymal cell fraction was resuspended in a solution with 9 ml ficoll and 1 ml percoll. One milliliter PBS was added carefully and spin at 4 °C for 15 min at 1400 × g in centrifuge. HSC cell fraction was obtained from the layer between PBS and the ficoll/percoll gradient solution. Kupffer cells were isolated using Optiprep38 (link). The nonparenchymal cells are resuspended in 20% Optiprep. HBSS and 11.5% Optiprep were layered on the cell suspension and centrifuged at 1811 × g for 17 m. Kupffer cell fraction was obtained from the layer between 20% Optiprep and 11.5% Optiprep.
Collagen coated plate
Collagen-coated plates are laboratory equipment designed for cell culture applications. The plates are coated with collagen, a naturally occurring extracellular matrix protein, to provide a suitable substrate for the attachment and growth of cells. These plates are intended to support the in vitro cultivation of various cell types.
Lab products found in correlation
19 protocols using collagen coated plate
Isolation of Liver Cell Subpopulations
The supernatant was centrifuged at 600 × g to obtain hepatic nonparenchymal cells. Primary HSCs were isolated by density-gradient centrifugation using ficoll paque-plus (GE Healthcare, Madison, WI) and percoll (GE Healthcare, Madison, WI, USA)41 (link). Non-parenchymal cell fraction was resuspended in a solution with 9 ml ficoll and 1 ml percoll. One milliliter PBS was added carefully and spin at 4 °C for 15 min at 1400 × g in centrifuge. HSC cell fraction was obtained from the layer between PBS and the ficoll/percoll gradient solution. Kupffer cells were isolated using Optiprep38 (link). The nonparenchymal cells are resuspended in 20% Optiprep. HBSS and 11.5% Optiprep were layered on the cell suspension and centrifuged at 1811 × g for 17 m. Kupffer cell fraction was obtained from the layer between 20% Optiprep and 11.5% Optiprep.
Isolation and Culture of Murine Hepatocytes
YAP inhibitory signals were introduced as following: Serum starvation (16 h); high density (100% confluence, 24 h); sorbitol (0.5 M, 1 h); cytochalasin D (1 μM, 1 h); latrunculin B (0.25 mg/ml, 1 h); and 2-DG (2-deoxyglucose; 25 mM, 1 h). LB-100 was pretreated in LATS1/2 KO cells at a concentration of 5 μM for 1 h and in wild-type cells at a concentration of 10 μM for 1 h.
Mammary Gland Organoid Self-Renewal Assay
Primary ErbB2-tumor cells were occasionally kept in short-term cultures in adhesion using DMEM/F12 (1:1, Lonza/GIBCO) supplemented with 10% FBS (HyClone, GE Healthcare Life Science), 10 mM HEPES (Sigma), 5 μg/mL insulin (Roche), 0.5 μg/mL hydrocortisone (Sigma), 10 ng/mL epidermal growth factor (EGF, Tebu-Bio), and 10 ng/mL cholera toxin (Sigma). In vitro treatments with Nut3 (Sigma, N6287), Adriamycin (Sigma, D1515) and 4-OHT (Sigma, H7904) were performed as indicated in the corresponding Results sections and Figures.
Isolation of Bovine, Porcine, and Chicken MSCs
Sebocyte and ORS Cell Viability Assay
Murine Mammary Organoid Culture
Isolation and Culture of Primary Hepatocytes and HSCs
Isolation and Differentiation of Murine Muscle Stem Cells
Neuronal Differentiation of SH-SY5Y Cells
Cryopreserved Mammary Epithelial Cell Isolation
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