Cells were seeded in 12-well plates (Nunc Delta Surface; Nunc, Wiesbaden, Germany). Cells were infected at an MOI of 0.03 in a total of 1 mL medium. At the indicated time points, supernatants were clarified by centrifugation and stored in aliquots at −80°C. Infected cells were scraped into 1 mL OptiMEM and subjected to a freeze-thaw cycle. After thawing, supernatants containing released particles were also clarified by centrifugation and stored in aliquots at −80°C. Cell-associated virus titers and titers of virus in supernatants were determined by TCID50 titration. Viral titers were analyzed 48 h after infection.
Nunc delta surface
Manufactured by Thermo Fisher Scientific
Sourced in Denmark
The Nunc Delta Surface is a specialized laboratory surface designed to optimize cell growth and attachment. It features a modified surface treatment that enhances the adhesion and proliferation of a variety of cell types. The Nunc Delta Surface is suitable for various cell culture applications, providing a reliable and consistent platform for researchers.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Labtek microscope dishes, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Lachrom elite, by Hitachi (1 mentions)
Beas 2b, by American Type Culture Collection (1 mentions)
Bronchial epithelial growth medium (begm), by Lonza (1 mentions)
Nci h292, by American Type Culture Collection (1 mentions)
Calu 3, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by Lonza (1 mentions)
Nci h441, by Thermo Fisher Scientific (1 mentions)
Nci h441, by American Type Culture Collection (1 mentions)
Beas 2b, by Lonza (1 mentions)
3 protocols using nunc delta surface
1
Virus Titer Determination Protocol
2
Culturing Glucose-Responsive MIN6 Cells
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MIN6 cells (Miyazaki et al., 1990 (link)) were cultured in a humidified atmosphere at 37°C and 8% CO2. The culture medium DMEM contained 4.5 g/L glucose and was supplemented with FBS (15%, gradient‐grade, Thermo Fisher Scientific, catalog# 16000‐044) and β‐mercaptoethanol (70 µM). The medium was sterile‐filtered (Millex GV, 0.22 µm) and used within 1 week after preparation. For imaging, MIN6 cells were seeded into 8‐well LabTek microscope dishes (155411 Thermo Scientific) or on 40 mm coverslips (Menzel Gläser). For in vitro assays, MIN6 cells were seeded on ∅ 60 mm or ∅ 35 mm dishes (Nunc delta surface, cat# 150288/cat# 153066; Roskilde, Denmark) to form pseudoislets within 5 days after seeding. MIN6 cells were used exclusively from passages 26 to 36.
3
Comparative analysis of immortalized lung cell lines
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Immortalized lung cell lines, Calu-3, BEAS2-B, NCI-H292, NCI-H441, and A549, were obtained from the American Type Culture Collection (ATCC) (Manassas, Virginia) in Nippon Boehringer Ingelheim and used for this study. These cell lines were seeded at a density of 1 × 10 5 cells/cm 2 on the 75 cm 2 of Nunc Delta Surface (Lot no. 12989; Nunc, Roslilde, Denmark). Calu-3 (passage number: 19-21) was cultured in EMEM (Lonza, Basel, Switzerland), BEAS2-B (passage number: 41-50) in BEGM (Lonza), NCI-H292 (passage number: 10-12), and NCI-H441 (passage number: 12-13) in RPMI1640 (Invitrogen, Carlsbad, California), and A549 (passage number: 80-90) in DMEM (Lonza), including 10% fetal bovine serum (Moregate, Bulimba, Australia), for 7 days at 37°C in 5% CO 2 , and the medium was exchanged every 2 days. All peptides were selected according to the in silico selection criteria reported previously 14 and synthesized by Thermoelectron Corporation (Sedanstrabe, Germany) at a peptide purity of more than 95%. The concentrations of peptide solutions were determined by quantitative amino acid analysis (Lachrom Elite, Hitachi, Tokyo, Japan). Other chemicals used were commercial products of an analytical grade.
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