Human lung epithelial A549 cells (Cat# 86012804-1VL, Sigma-Aldrich) and human distal lung epithelial H441 cells (Cat# ATCC-CRM-HTB-174D, LGC) were cultured in F-12K Nut Mix medium (Kaighn’s modification, Gibco) supplemented with 10% (v/v) artificial fetal calf serum (FCS) (HyClone FetalClone III serum, Cytiva). T7 mouse type-II alveolar epithelial cells (Cat# 07021402, ECACC) were cultured in F-12K Nut Mix medium supplemented with 5% (v/v) artificial FCS and with 0.5% (v/v) Insulin-Transferrin-Selenium (Thermo Fisher Scientific). BEAS-2B cells (Cat# CRL-9609, ATCC) were cultured in LHC-9 serum free medium (Thermo Fisher Scientific) in flasks precoated with LHC-9 medium supplemented with 0.01 mg/mL bovine fibronectin (Thermo Fisher Scientific), 0.03 mg/mL bovine collagen type I (Sigma-Aldrich) and 0.01 mg/mL BSA (Sigma-Aldrich). HepG2 cells (Cat# ACC 180, DSMZ) were cultured in RPMI-1640 medium supplemented with 10% (v/v) artificial FCS. A549 cells stably expressing Cas9 (Cat# SL504, GeneCopoiea) were cultured as mentioned above for A549 cells, but with addition of 800 μg/mL hygromycin (InvivoGen) as selection marker. The primary human airway epithelial (pAEC) cells (Cat# CC-2547, Lonza) were cultured in SAGM Small Airway Epithelial Cell Growth Medium BulletKit (Lonza). All cells were cultivated at 37°C and 5% (v/v) CO2.
Lhc 9 serum free medium
Manufactured by Thermo Fisher Scientific
Sourced in France, United States
The LHC-9 serum-free medium is a cell culture media formulation designed to support the growth and maintenance of a variety of cell lines in the absence of serum. It provides the necessary nutrients and growth factors for optimal cell proliferation and viability.
Automatically generated - may contain errors
Lab products found in correlation
Insulin transferrin selenium, by Thermo Fisher Scientific (1 mentions)
Beas 2b cell, by American Type Culture Collection (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Bovine fibronectin, by Thermo Fisher Scientific (1 mentions)
Hygromycin, by InvivoGen (1 mentions)
Hyclone fetalclone 3 serum, by Cytiva (1 mentions)
Bovine collagen type 1, by Merck Group (1 mentions)
Sagm small airway epithelial cell growth medium bulletkit, by Lonza (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Human fibronectin, by Thermo Fisher Scientific (1 mentions)
Lhc basal medium, by Thermo Fisher Scientific (1 mentions)
Begm medium, by Lonza (1 mentions)
Trypsin edta 0.25, by Thermo Fisher Scientific (1 mentions)
F 12k medium, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
A549 cells ccl 185, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Beas 2b cells, by Thermo Fisher Scientific (1 mentions)
Sterile tissue culture flasks, by Thermo Fisher Scientific (1 mentions)
Mycoalert plus mycoplasma detection kit, by Lonza (1 mentions)
3 protocols using lhc 9 serum free medium
1
Cultivation of Diverse Lung Epithelial Cell Lines
2
Characterization of BEAS-2B Bronchial Epithelial Cells
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The BEAS-2B cell line is an immortalized but non-tumorigenic human cell line established from normal human bronchial epithelium obtained from a healthy individual [50 (link)]. This cell line has been widely used to determine various chemical, biological, and particle agents with potential pulmonary toxicity [9 (link),42 (link),51 (link)]. Cells were obtained from the American Type Culture Collection (CRL#9609, LGC Standards Sarl, Molsheim, France). They were cultured in culture flasks precoated with LHC basal medium supplemented with BSA (0.01 mg/mL), human fibronectin (0.01 mg/mL), and collagen (0.03 mg/mL) (Thermo Fisher Scientific, Illkirch, France). They were maintained in LHC-9 serum-free medium (Thermo Fisher Scientific, Illkirch, France) and BEGMTM Medium (Lonza; Basel, Switzerland) in a humidified atmosphere of 5% CO2 at 37 °C. Before confluence, passage was performed twice a week using trypsin (0.25%)-EDTA (2.6 mM) (Thermo Fisher Scientific, Illkirch, France).
3
Cell Culture Protocols for A549 and BEAS-2B
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The A549 cells (CCL185) and BEAS-2B cells (CRL-9609) were obtained from American Type Culture Collection (ATCC, LGC Standards, UK). The A549 cells were cultured as a monolayer in an F12K medium (Gibco, Invitrogen, Carlsbad, CA, USA) supplemented with 10% foetal bovine serum (FBS, Gibco) and 1% antibiotic-antimycotic (Sigma-Aldrich) in sterile tissue culture flasks (Nunc, Roskilde, Denmark). BEAS-2B cells were cultured in the LHC-9 serum-free medium (Thermo Fisher Scientific MA, USA) containing 1% antibiotic-antimycotic (Sigma-Aldrich). Both cell lines were grown at 37 °C and in a humidified atmosphere (5% CO2). The cells were passaged before reaching 90% confluence. To detach cells from the culture plates, 0.25% trypsin-EDTA (Gibco) was used. Additionally, cells were screened for Mycoplasma sp. infection using MycoAlert™ PLUS Mycoplasma Detection Kit (Lonza, Walkersville, Inc.).
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