Primary human hepatic stellate cells (pHSCs), primary human liver sinusoidal endothelial cells (pLSECs) and primary human kupffer cells were obtained from Sciencell (Sciencell, CA). pHSCs were cultured on poly(L-lysine) coated plates in stellate cell medium (StECM, Sciencell) supplemented with stellate cells growth supplements (SteCGS), 2% FBS and antibiotics (Penicillin and streptomycin). pLSECs were cultured on fibronectin-coated plates in endothelial cell medium (ECM, Sciencell) supplemented with endothelial cells growth supplements (ECGS), 5% FBS and antibiotics (Penicillin and streptomycin). Primary human kupffer cells were cultured on poly(L-lysine) coated plates in macrophage medium (MM, Sciencell) supplemented with macrophage growth supplements (MaGS), 5% FBS and antibiotics (Penicillin and streptomycin).
Stecm
Manufactured by ScienCell
Sourced in United States
SteCM is a specialized cell culture medium designed to support the growth and maintenance of stem cells. It provides the necessary nutrients, growth factors, and other components required for the optimal expansion and preservation of stem cell characteristics.
Automatically generated - may contain errors
Lab products found in correlation
Foetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Poly l lysine (pll), by Merck Group (2 mentions)
Stellate cell growth supplement, by ScienCell (2 mentions)
Fetal bovine serum (fbs), by ScienCell (2 mentions)
Phscs, by ScienCell (1 mentions)
Endothelial cell medium (ecm), by ScienCell (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Lonza (1 mentions)
Rpmi 1640 medium, by Lonza (1 mentions)
Primary human hscs, by ScienCell (1 mentions)
Penicillin streptomycin, by ScienCell (1 mentions)
Penicillin, by ScienCell (1 mentions)
Streptomycin, by ScienCell (1 mentions)
Hhstec, by ScienCell (1 mentions)
9 protocols using stecm
1
Culturing Primary Liver Cell Types
2
Primary Hepatic Stellate Cell Cultivation
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Primary Human hepatic stellate cells (HSCs) were purchased from Zen-Bio (Durham, NC, USA) and were grown in stellate cell growth medium SteCM (ScienCell, Carlsbad, CA, USA) according to the manufacturer’s instructions. HepG2 and THP1 cells were purchased from ATCC (Manassas, VA, USA) and ECACC (Sigma, St Louis, MO, USA), respectively, and were cultured in Dulbecco’s modified Eagle’s medium and RPMI-1640 medium (Lonza, Verviers, Belgium), supplemented with 2 mM L-glutamine, 10% fetal bovine serum (FBS, Lonza) and antibiotics (50 U ml−1 Penicillin and 50 μg ml−1 streptomycin, Sigma) respectively. No cell line used in this paper is listed in the database of commonly misidentified cell lines that is maintained by the International Cell Line Authentication Committee (http://iclac.org/databases/cross-contaminations/ ). All cell lines were tested negative for mycoplasma contamination.
3
Hepatic Stellate Cell Collagen Response
4
Activated HSCs Cultured with hAMSC-CM
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Approximately 5 × 104 HSCs were seeded on 12-well plastic plates, on which the cells were automatically activated and proliferated [16 (link)]. They were cultured in 2 mL of stellate cell medium (SteCM; ScienCell, Carlsbad, CA, USA) supplemented with 2% FBS (ScienCell) and stellate cell growth supplement (SteCGS; ScienCell) for 48 h. Then, the HSCs were washed thrice using HBSS (−) and were cultured for 48 h with SM or hAMSC-CM. In addition, hAMSC-CM was mixed separately with the SM in two concentrations: 50% and 25% (v/v) of hAMSC-CM. These different concentrations of hAMSC-CM were also used for culturing washed HSCs for 48 h as described above.
5
Expansion of Primary Human HSCs
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Primary human HSCs were purchased from ScienCell Research Laboratories (ScienCell, Carlsbad, CA, USA). Cells were expanded by using stellate cell medium (SteCM) (ScienCell), in a humidified incubator under 5% CO2 and 95% air at 37 °C. The third‐passaged cells were used for the following experiments.
6
Cultivation of Primary Human Pancreatic Stellate Cells
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Primary human pancreatic stellate cells (PSCs; ScienCell, Carlsbad, CA, USA; Cat #3830, Lot #14289) were isolated from human pancreas healthy samples and were maintained in Stellate cell medium (SteCM, Cat #5301, ScienCell, Carlsbad, CA, USA) supplemented with stellate cell growth supplement(ScienCell, Carlsbad, CA, USA Cat #5352), 2% fetal bovine serum (ScienCell, Carlsbad, CA, USA; Cat #0010) and 1% Penicillin-streptomycin (ScienCell, Carlsbad, CA, USA Cat #0503), in an incubator at 37 °C and with an atmosphere of 5% CO2. Unless otherwise specified, PSCs were serum-starved overnight prior to 24 h stimulation with different doses of various recombinant proteins, as outlined in the main text and/or figure legends. All experiments were carried out at low cell passage (
7
Hepatic Stellate Cell Collagen Response
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Primary human hepatic stellate cells (HSC) were purchased (ScienCell, 5300) and cultured in stellate cell medium (SteCM, ScienCell, 5301) on Poly-L-Lysine (Sigma, P4832) coated T75 tissue culture flasks, according to the suppliers protocol. All experiments were performed using cells between passage 3 and 5. For assessment of fibrillar collagen gene expression, HSC were plated at 75,000 cells per well in 24 well-plates (Costar, 3524) in HSC media consisting of DMEM (Gibco, 21969-035) supplemented with 20 µM HEPES (Sigma, H3375,), 2 mM L-Glutamine (Gibco, 25030-024), 1% Penicillin Streptomycin (Gibco, 15140-122, Gibco) and 2% Foetal Bovine Serum (FBS, Gibco, 10270). HSC were serum starved overnight (in HSC media without FBS), washed with PBS, then 250μl of conditioned media from primary human macrophage subpopulations added for 24 hours. HSC were harvested for RNA.
8
Culturing and Cryopreservation of Primary Hepatic Stellate Cells
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Primary activated human hepatic stellate cells (HSC) obtained from ScienCell (San Diego, CA, USA) have been well characterized [18 (link)]. HSC were cultured for 2–4 passages in defined Stellate Cell Medium (SteCM, ScienCell) supplemented with 2% fetal bovine serum, 5 ml stellate cell growth supplement, 10 U/ml penicillin and 10 μg/ml streptomycin (all ingredients obtained from ScienCell) at 37°C with 5% CO2. Then, cells were cryopreserved until further use.
Two days before HSC were used in an experiment, the cells were thawed and cultured in SteCM medium. Then cells were harvested, washed, checked for viability using trypan blue, and used in the respective experiments.
Two days before HSC were used in an experiment, the cells were thawed and cultured in SteCM medium. Then cells were harvested, washed, checked for viability using trypan blue, and used in the respective experiments.
9
Culturing Human Hepatic Stellate Cells
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HHSteCs were purchased from ScienCell Research Laboratories (San Diego, CA) and cultured in complete stellate cell medium (SteCM, ScienCell Research Laboratories) supplemented with 2% fetal bovine serum (FBS, ScienCell Research Laboratories), 1% stellate cell growth supplement (S, ScienCell Research Laboratories), and 1% penicillin-streptomycin (P/S, ScienCell Research Laboratories) at 37 °C in humidified air containing 5% CO2. HHSteCs were passaged when sub-confluent and were used between passages 3–10 for all experiments. See Supplementary Materials for details.
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