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Uncoated plastic dishes

Manufactured by BD
Sourced in United States

Uncoated plastic dishes are sterile, disposable laboratory containers used for various applications in cell culture, microbiological studies, and other scientific research. They provide a flat, smooth surface for the growth and observation of cells, microorganisms, or other samples. The dishes are made of high-quality, non-treated plastic materials that are suitable for a range of laboratory procedures.

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5 protocols using uncoated plastic dishes

1

Isolation and Culture of Primary Mouse HSC

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The pronase and collagenase digestion method was used to isolate primary mouse HSC from C57BL/6J mice, as previously described [23 (link)]. Primary mouse HSC were plated on uncoated plastic dishes (BD Falcon, Franklin Lakes, NJ, USA) for spontaneous activation [4 (link)] and maintained in low-glucose Dulbecco’s Modified Eagle Medium supplemented with 10% fetal bovine serum, 4 mM L-glutamine, penicillin (100 U/mL), and streptomycin (100 μg/mL) as previously described [23 (link),25 (link)]. The cells were cultured at 37 °C under 5% CO2. Cell culture supplies were purchased from HyClone (Thermo Scientific, Logan, UT, USA). The cells at day 1 and day 7 after isolation represent qHSC and aHSC, respectively.
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2

Primary Mouse and Human HSC Isolation and Culture

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Primary mouse HSCs were isolated from C57BL/6J mice (Jackson Laboratory, Bar Harbor, ME, USA) using a pronase/collagenase digestion method and cultured on uncoated plastic dishes (BD Falcon, Franklin Lakes, NJ, USA) for activation (Friedman, 2008 (link)) as previously described (Yang et al., 2016a (link), Yang et al., 2015a (link)). Primary mouse HSCs were cultured for 1 day or 7 days to represent qHSCs or aHSCs, respectively. LX-2 cells, a human HSC cell line kindly provided by Dr. Scott Friedman at the Icahn School of Medicine at Mount Sinai (New York, NY, USA), were cultured as previously described (Yang et al., 2015a (link)). Primary human HSCs (Zen-Bio, Research Triangle Park, NC, USA) were cultured in a Human Stellate Growth Medium (Zen-Bio) on collagen-coated plates (Corning life sciences, Oneonta, NY, USA). ASTX, a generous gift from Fuji Chemical Industry Co., Ltd. (Toyama, Japan), was treated as previously described (Yang et al., 2015a (link)). Primary mouse HSCs were treated with 25 μM ASTX or with 2, 5, or 10 mM sodium L-lactate (Sigma, St. Louis, MO, USA) for 5 days during activation with daily replenishment. All cells were maintained in a 37°C humidified cell culture chamber with 5% CO2. Cell culture supplies were purchased from HyClone (Thermo Scientific, Logan, UT, USA) unless stated otherwise.
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3

Isolation and Culture of Hematopoietic Stem Cells

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HSCs were isolated from WT and Cygb−/− mice using the pronase-collagenase digestion method as described previously66 (link) and were cultured on uncoated-plastic dishes (BD Falcon, Franklin Lake, NY, USA) or glass chamber slides (Thermo Fisher Scientific, Waltham, MA, USA) in DMEM (Sigma-Aldrich) supplemented with 10% FBS (Invitrogen, Carlsbad, CA, USA) and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin) at 37 °C in a 5% CO2/95% room air. Mouse hepatoma Hepa 1–6 cells (CRL-1830) obtained from American Type Culture Collection (Manassas, VA, USA) were maintained on uncoated-plastic culture plates (BD Falcon) in DMEM supplemented with 10% FBS and antibiotic.
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4

Isolation and Activation of Mouse Hepatic Stellate Cells

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Primary mouse HSC were isolated from the liver of C57BL/6J mice using the pronase and collagenase digestion method as described in detail previously [20 (link)]. To activate the cells, HSC were cultured on uncoated plastic dishes (BD Falcon, Franklin Lakes, NJ, USA) [21 (link)] in low-glucose Dulbecco’s modified Eagle medium supplemented with fetal bovine serum (10%), L-glutamine (4 mM), penicillin (100 U/mL), and streptomycin (100 μg/mL) as previously described [20 (link),22 (link)]. The cells were maintained at 37 °C under 5% CO2. Cell culture supplies were purchased from HyClone (Thermo Scientific, Logan, UT, USA). The cells at day 1 and day 7 after isolation were considered as qHSC and aHSC, respectively.
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5

Isolation and culture of hepatic stellate cells

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HSCs were isolated from WT (HSC CygbÀwild ) and Cygb À/À (HSC CygbÀnull ) mice using the pronase-collagenase digestion method, as previously described, 22 and were cultured on uncoated plastic dishes (BD Falcon, Franklin Lake, NY) or glass chamber slides (Thermo Fisher Scientific, Waltham, MA) in Dulbecco's modified Eagle's medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA) and antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin). HSC CygbÀwild and HSC CygbÀnull cells were harvested at days 1, 4, and 7 for RNA, protein extractions, or for immunofluorescence, Oil Red O staining.
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