Hepatocyte culture media

Manufactured by Lonza

Sourced in United States

Hepatocyte Culture Media is a specialized cell culture medium designed to support the growth and maintenance of primary human hepatocytes. It provides the necessary nutrients, growth factors, and biochemical components to facilitate the in vitro culture and expansion of these liver-derived cells.

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Lab products found in correlation

Oncostatin m, by R&D Systems (2 mentions) Activin a, by R&D Systems (2 mentions) Hepa1 6 cells, by American Type Culture Collection (1 mentions) Hcm singlequot, by Lonza (1 mentions) Epidermal growth factor (egf), by Lonza (1 mentions) Geltrex, by Thermo Fisher Scientific (1 mentions) Rpmi b27 without insulin, by Thermo Fisher Scientific (1 mentions) Rpmi b27, by R&D Systems (1 mentions) Bone morphogenetic protein 4 (bmp4), by R&D Systems (1 mentions) Fibroblast growth factor 2 (fgf2), by R&D Systems (1 mentions) Mtesr1, by STEMCELL (1 mentions) Hepatocyte growth factor (hgf), by R&D Systems (1 mentions) Fibroblast growth factor 2 (fgf2), by Thermo Fisher Scientific (1 mentions) Bone morphogenetic protein 4 (bmp4), by Thermo Fisher Scientific (1 mentions) Matrigel, by Thermo Fisher Scientific (1 mentions) Rpmi b27, by Thermo Fisher Scientific (1 mentions) Matrigel, by R&D Systems (1 mentions) Accutase, by Merck Group (1 mentions) Hepatocyte growth factor (hgf), by Thermo Fisher Scientific (1 mentions)

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Culture media

5 protocols using hepatocyte culture media

Hepa1-6 Cell Culture and Primary Hepatocyte Isolation

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Hepa1-6 cells were obtained from the American Type Culture Collection (Manassas). Hepa1-6 cells were maintained in DMEM containing 10% heat-inactivated FBS and antibiotics (100 U/mL penicillin, 100 μg/mL streptomycin). Cells were cultured at 37°C under an atmosphere of 95% air and 5% CO₂.
Isolation of primary hepatocytes was described previously (Ariyoshi et al. 2017 (link)). Briefly, bone marrow cells were cultured in 1 mL Hepatocyte Culture Media (Lonza Walkersville, Walkersville, MD, USA) supplemented with 5% FBS for 6 h in a 37°C incubator with 5% CO₂. One part conditioned medium was added to four parts of liver cell culture. Livers were minced to the level of 2 mm³. The pieces of liver were incubated for 30 min at 37°C with collagenase-Yakult (100 U/mL, Yakult Pharmaceutical, Tokyo, Japan). After collagenase treatment, the supernatant was decanted, and 5 mL of HGM supplemented with 10% FBS was added; the cells were then shaken at 160 rpm for 30 min at room temperature. After centrifugation at 50 × g for 2 min, the supernatant was removed, and 5 mL fresh HGM was added.

Islam M.N., Mita Y., Maruyama K., Tanida R., Zhang W., Sakoda H, & Nakazato M. (2019). Liver-expressed antimicrobial peptide 2 antagonizes the effect of ghrelin in rodents. The Journal of Endocrinology, 244(1), 13-23.

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Primary Hepatic Cell Culture Protocol

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Primary human hepatic (PHH) cells were purchased and cultured in hepatocyte culture media (LONZA, Walkersville) at 37°C with 5% CO₂ according to the manufacturer's recommendations. The hepatocarcinoma cell (HCC) lines, HepG2, HuH7, HLF, and PLC/PRF/5 and the control cell line HEK293 obtained from ATCC or JCRB cell bank were cultured in Dulbecco's modified Eagle's medium or RPMI1640 medium supplemented with antibiotics and 10% fetal bovine serum. HAK1A and HAK1B (24 (link)) kindly gifted HCC lines were cultured in RPMI1640 medium with antibiotics and 10% fetal bovine serum.

Angata K., Sawaki H., Tsujikawa S., Ocho M., Togayachi A, & Narimatsu H. (2020). Glycogene Expression Profiling of Hepatic Cells by RNA-Seq Analysis for Glyco-Biomarker Identification. Frontiers in Oncology, 10, 1224.

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Evaluating HDL Nanoparticles in Hepatocytes

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Primary human hepatocytes were obtained from Lonza and cultured in Hepatocyte Culture Media (HCM), consisting of Hepatocyte Basal Media (Lonza) supplemented with Lonza’s HCM SingleQuot, as described previously (26 ). Briefly, cells were plated at a density of 1 × 10⁵ cells/ml and allowed to attach overnight prior to treatment with HDL NPs. Cells were treated with 20 nM or 50 nM HDL NPs, 50 nM human HDL or PBS for 72 or 120 h. Lysates were prepared using the mammalian protein extraction reagent (MPER) cell lysis buffer, and Western blotting analyses for GPX4 and β-actin conducted as described above. For the C11-BODIPY assay, the primary human hepatocytes were treated for 72 h prior to addition of the C11-BODIPY reagent and subsequent analysis by flow cytometry.

Rink J.S., Lin A.Y., McMahon K.M., Calvert A.E., Yang S., Taxter T., Moreira J., Chadburn A., Behdad A., Karmali R., Thaxton C.S, & Gordon L.I. (2020). Targeted reduction of cholesterol uptake in cholesterol-addicted lymphoma cells blocks turnover of oxidized lipids to cause ferroptosis. The Journal of Biological Chemistry, 296, 100100.

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Differentiation of hPSCs into Hepatocytes

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hPSCs were differentiated as discussed previously (Si-Tayeb et al. 2010 (link); Mallanna and Duncan 2013 ; Yanagihara et al. 2016 (link)). Briefly, hPSCs were harvested using Accutase and plated at 600,000 cells per well in 24-well plates precoated with 300 µl/well Geltrex (Thermo Fisher Scientific, Waltham, MA). Approximately 24 h after seeding the cells with mTeSR1 (Stemcell Technologies), when the cells were 85–95% confluent, differentiation was initiated by culture for 5 d with 50 ng/ml Activin A (R&D Systems) in RPMI/B27 (without insulin) supplement (Invitrogen) under ambient oxygen/5% CO₂. In addition, we included 10 ng/ml BMP4 (R&D Systems) and 20 ng/ml FGF2 (R&D Systems) for the first 2 d. Then, the cells were cultured for 5 d with 20 ng/ml BMP4 (R&D Systems)/10 ng/ml FGF-2 (R&D Systems) in RPMI/B27 (containing insulin) under 4% O₂/5% CO₂, then for 5 d with 20 ng/ml HGF (R&D Systems) in RPMI/B27 (containing insulin) under 4% O₂/5% CO₂, and finally for 5 d with 20 ng/ml Oncostatin-M (R&D Systems) in Hepatocyte Culture Media (Lonza) supplemented with SingleQuots (without EGF) in ambient oxygen/5% CO₂.

Yanagihara K., Hayashi Y., Liu Y., Yamaguchi T., Hemmi Y., Kokunugi M., Yamada K.U., Fukumoto K., Suga M., Terada S., Nikawa H., Kawabata K, & Furue M. (2024). Trisomy 12 compromises the mesendodermal differentiation propensity of human pluripotent stem cells. In Vitro Cellular & Developmental Biology. Animal, 60(5), 521-534.

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Differentiation of Pluripotent Cells into Hepatocytes

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Pluripotent cells were differentiated as discussed previously
[14 (link),18 ]. Briefly, pluripotent cells cultured on E-cadherin-IgG Fc were harvested using Accutase (Millipore) and plated onto 6-well tissue culture-treated plates pre-coated with 2 mg/ml Matrigel (Geltrex; Invitrogen). Typically, one 100 mm dish of cells cultured on E-cadherin-IgG Fc provided enough cells for 2 wells of a 6-well plate. Approximately 24 hours after seeding the cells onto Matrigel, when the cells were 85-95% confluent, differentiation was initiated by culture for 5 days with 50 ng/ml Activin A (R&D Systems) in RPMI/B27 (without Insulin) supplement (Invitrogen) under ambient oxygen/5%CO₂. In addition, we included 10 ng/ml BMP4 (Peprotech) and 20 ng/ml FGF-2 (Invitrogen) for the first 2 days. This resulted in reproducible differentiation into definitive endoderm at efficiencies of greater than 80%. Cells were cultured for 5 days with 20 ng/ml BMP4 (Peprotech)/10 ng/ml FGF-2 (Invitrogen) in RPMI/B27 (containing Insulin) under 4%O₂/5%CO₂, then 5 days with 20 ng/ml HGF (Peprotech) in RPMI/B27 (containing Insulin) under 4%O₂/5%CO₂, and finally for 5 days with 20 ng/ml Oncostatin-M (R&D Systems) in Hepatocyte Culture Media (Lonza) supplemented with SingleQuots (without EGF) in ambient oxygen/5% CO₂.

Noto F.K., Determan M.R., Cai J., Cayo M.A., Mallanna S.K, & Duncan S.A. (2014). Aneuploidy is permissive for hepatocyte-like cell differentiation from human induced pluripotent stem cells. BMC Research Notes, 7, 437.

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