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Cc 3118

Manufactured by Lonza
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The CC-3118 is a laboratory equipment product from Lonza. It is a compact, centrifuge designed for general laboratory applications. The core function of the CC-3118 is to separate components of a liquid mixture based on their density differences through the application of centrifugal force.

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Lab products found in correlation

Pneumacult ali medium, by STEMCELL (3 mentions) Recombinant murine il 13, by Thermo Fisher Scientific (2 mentions) Deferoxamine mesylate, by Selleck Chemicals (1 mentions) Necrosulfonamide, by Selleck Chemicals (1 mentions) Z vad fmk, by Selleck Chemicals (1 mentions) Pcs 301 010, by American Type Culture Collection (1 mentions) Pcs 300 030, by American Type Culture Collection (1 mentions) Pcs 300 040, by American Type Culture Collection (1 mentions) Evos fl microscope, by Thermo Fisher Scientific (1 mentions) Cholera toxin, by Merck Group (1 mentions)

8 protocols using cc 3118

1

Culturing A549 and Small Airway Cells

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A549 cells (American Type Culture Collection, #CCL-185) were cultured in DMEM supplemented with 10% heat-inactivated fetal bovine serum (FBS). The cells were passaged <20 times. Small airway epithelial cells (American Type Culture Collection, #PCS-301-010) were cultured in small airway epithelial growth media (Lonza #CC-3118) and passaged < 7 times. For studies using cell death inhibitors, deferoxamine mesylate (Selleckchem #S5742), necrosulfonamide (Selleckchem #S8251), and Z-VAD-FMK (Selleckchem #S7023) were dissolved in DMSO, and cells were treated at indicated concentrations
Sauler M., McDonough J.E., Adams T.S., Kothapalli N., Barnthaler T., Werder R.B., Schupp J.C., Nouws J., Robertson M.J., Coarfa C., Yang T., Chioccioli M., Omote N., Cosme C J.r., Poli S., Ayaub E.A., Chu S.G., Jensen K.H., Gomez J.L., Britto C.J., Raredon M.S., Niklason L.E., Wilson A.A., Timshel P.N., Kaminski N, & Rosas I.O. (2022). Characterization of the COPD alveolar niche using single-cell RNA sequencing. Nature Communications, 13, 494.
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2

Immortalization of Human Small-Airway Epithelial Cells

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Human small-airway epithelial cells (hSAECs) from a cadaveric donor were immortalized using human telomerase/CDK4, as previously described [20 (link),21 (link)]. These non-oncogenic, telomerase-immortalized cells maintain genomic and proteomic signatures representative of primary SAECs over many population doublings [21 (link)] and manifest characteristic cell-state transitions typical of primary hSAECs [20 (link),22 (link),23 (link)]. hSAECs were grown in small-airway epithelial cell growth medium (Lonza, cc-3118) in a humidified atmosphere of 5% CO2. For validation experiments, primary small-airway epithelial cells (pSAECs) were obtained from cadaveric donors (ATCC, PCS-301-010). Cells were grown in airway epithelial cell basal medium (ATCC, PCS-300-030) supplemented with a bronchial epithelial cell growth kit (PCS-300-040).
Xu X., Mann M., Qiao D, & Brasier A.R. (2021). Alternative mRNA Processing of Innate Response Pathways in Respiratory Syncytial Virus (RSV) Infection. Viruses, 13(2), 218.
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3

Airway Basal Stem Cell Differentiation

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Tracheal cells were harvested and cultured as previously described (Mou et al., 2016 (link)). Briefly, cells were cultured and expanded in complete SAGM (small airway epithelial cell growth medium; Lonza, CC-3118) containing a TGF-β/BMP4/WNT antagonist cocktail and 5 μM ROCK inhibitor Y-27632 (Selleckbio, S1049). To generate ALI cultures, airway basal stem cells were seeded onto transwell membranes and differentiated using PneumaCult-ALI Medium (StemCell, 05001). Air-liquid cultures were fully differentiated after 14 days as demonstrated by the formation of air-liquid interface and the beating of cilia.
Shah V.S., Hou J., Vinarsky V., Xu J., Surve M.V., Lin C.P, & Rajagopal J. (2023). Autofluorescence imaging permits label-free cell type assignment and reveals the dynamic formation of airway secretory cell associated antigen passages (SAPs). eLife, 12, e84375.
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4

Mouse Lung Epithelial Cell Culture Protocol

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Mouse Lung Epithelial (MLE12) cells were obtained from ATCC and grown in HITES media as formulated by ATCC. Tunicamycin (Tun) was purchased from Millipore. STF-083010, KIRA7 and KIRA8 were synthesized in house. Mouse primary AECII cells were isolated as described [42 (link)]. Cells were grown in SAGM media (Lonza CC-3118) on fibronectin coated plates.
Thamsen M., Ghosh R., Auyeung V.C., Brumwell A., Chapman H.A., Backes B.J., Perara G., Maly D.J., Sheppard D, & Papa F.R. (2019). Small molecule inhibition of IRE1α kinase/RNase has anti-fibrotic effects in the lung. PLoS ONE, 14(1), e0209824.
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5

Clonal Alveolar Organoid Assay

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Clonal alveolar organoid assays were performed as described previously with some modifications from the original protocol17 (link),37 (link),81 (link). In brief, 5 × 103 AT2 cells were isolated as described above and mixed with 5 x104 lung PDGFRα+ isolated cells from adult wild type mice as previously described19 (link) for mouse or MRC5 cells (ATCC CCL-171, tested negative for mycobacterial contamination, at no greater than passage 7) for human cultures. For the first two days of culture, ROCK inhibitor Y27632 (Sigma-Aldrich catalog #Y0503) was added to the media (SAGM; Lonza catalog #CC-3118). After two days of culture, Y276632 was removed and ligand treatments of organoids were performed using the following reagents at the indicated concentrations: ANA-12 0.02μg/mL (Alomone Labs catalog #A-215), recombinant human BDNF 0.05μg/mL (Alomone Labs catalog #B-250) and recombinant murine FGF7 0.01μg/mL (R&D Systems catalog #5028-KG). DMSO was used as a control for the ANA-12 and 2% BSA was used as a control for the BDNF and FGF7. Media was changed every 48 hours, and fresh ligands were included at each media change. After 21 days (or 28 days where indicated) organoids were imaged using an Olympus MVX10 microscope or EVOS FL microscope (Life Technologies).
Paris A.J., Hayer K.E., Oved J.H., Avgousti D.C., Toulmin S.A., Zepp J.A., Zacharias W.J., Katzen J.B., Basil M.C., Kremp M.M., Slamowitz A.R., Jayachandran S., Sivakumar A., Dai N., Wang P., Frank D.B., Eisenlohr L.C., Cantu E I.I.I., Beers M.F., Weitzman M.D., Morrisey E.E, & Worthen G.S. (2020). STAT3-BDNF-TrkB signaling promotes alveolar epithelial regeneration after lung injury. Nature cell biology, 22(10), 1197-1210.
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6

Airway Basal Stem Cell Differentiation

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Cells were cultured and expanded in complete SAGM (small airway epithelial cell growth medium; Lonza, CC-3118) containing TGF-β/BMP4/WNT antagonist cocktails and 5 μM Rock inhibitor Y-27632 (Selleckbio, S1049). To initiate air–liquid interface (ALI) cultures, airway basal stem cells were dissociated from mouse tracheas and seeded onto transwell membranes. After reaching confluence, media was removed from the upper chamber. Mucocilliary differentiation was performed with PneumaCult-ALI Medium (StemCell, 05001). Differentiation of airway basal stem cells on an air–liquid interface was followed by directly visualizing beating cilia in real time after 10–14 days.
Once air-liquid cultures were fully differentiated, as indicated by beating cilia, treatment cultures were supplemented with 25ng/mL of recombinant murine IL-13 (Peprotech-stock diluted in water and used fresh) diluted in PneumaCult-ALI Medium, while control cultures received an equal volume of water for 72 hours. After treatment, whole ALI wells were fixed in 4% PFA, immunostained in whole mount using the same buffers and imaged with a confocal microscope as described above.
Montoro D.T., Haber A.L., Biton M., Vinarsky V., Lin B., Birket S., Yuan F., Chen S., Leung H.M., Villoria J., Rogel N., Burgin G., Tsankov A., Waghray A., Slyper M., Waldmann J., Nguyen L., Dionne D., Rozenblatt-Rosen O., Tata P.R., Mou H., Shivaraju M., Bihler H., Mense M., Tearney G.J., Rowe S.M., Engelhardt J.F., Regev A, & Rajagopal J. (2018). A revised airway epithelial hierarchy includes CFTR-expressing ionocytes. Nature, 560(7718), 319-324.
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7

Alveolar Organoid Culture Optimization

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Alveolar organoids were cultured as described previously29 (link),31 (link), with the following modifications. Briefly, 5 × 103 AT2s sorted from either SPCΔAb1 and Ab1fl/fl mice were co-cultured with 5 × 104 Pdgfrα+ lung fibroblasts sorted from WT C57Bl/6 mice. Cells were suspended in 90μL of a 1:1 mixture of Matrigel:modified SAGM media (CC-3118, Lonza). SAGM media is modified by adding 0.1 μg/mL cholera toxin (C8052, Sigma) and the following SAGM BulletKit components: 10 μg/mL insulin, 5 μg/mL transferrin, 25 ng/mL EGF, 30 μg/mL bovine pituitary extract, 0.01 μM retinoic acid, and 5% FBS. The 1:1 mixture containing cells was placed in a cell culture insert (353095, Corning); after solidification of the matrix at 37 °C, the insert was placed inside a well of 24-well plate containing modified SAGM media. For the first two days of culture, 10 μM ROCK inhibitor Y27632 (Y0503, Sigma) was added to the media. The media was changed every 48 h, and after 21 days organoids were imaged using an EVOS FL Auto using built-in EVOS FL Auto software. Organoid numbers were quantified in Fiji using the Cell Counter plugin.
Toulmin S.A., Bhadiadra C., Paris A.J., Lin J.H., Katzen J., Basil M.C., Morrisey E.E., Worthen G.S, & Eisenlohr L.C. (2021). Type II alveolar cell MHCII improves respiratory viral disease outcomes while exhibiting limited antigen presentation. Nature Communications, 12, 3993.
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8

Airway Basal Stem Cell Differentiation

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Cells were cultured and expanded in complete SAGM (small airway epithelial cell growth medium; Lonza, CC-3118) containing TGF-β/BMP4/WNT antagonist cocktails and 5 μM Rock inhibitor Y-27632 (Selleckbio, S1049). To initiate air–liquid interface (ALI) cultures, airway basal stem cells were dissociated from mouse tracheas and seeded onto transwell membranes. After reaching confluence, media was removed from the upper chamber. Mucocilliary differentiation was performed with PneumaCult-ALI Medium (StemCell, 05001). Differentiation of airway basal stem cells on an air–liquid interface was followed by directly visualizing beating cilia in real time after 10–14 days.
Once air-liquid cultures were fully differentiated, as indicated by beating cilia, treatment cultures were supplemented with 25ng/mL of recombinant murine IL-13 (Peprotech-stock diluted in water and used fresh) diluted in PneumaCult-ALI Medium, while control cultures received an equal volume of water for 72 hours. After treatment, whole ALI wells were fixed in 4% PFA, immunostained in whole mount using the same buffers and imaged with a confocal microscope as described above.
Montoro D.T., Haber A.L., Biton M., Vinarsky V., Lin B., Birket S., Yuan F., Chen S., Leung H.M., Villoria J., Rogel N., Burgin G., Tsankov A., Waghray A., Slyper M., Waldmann J., Nguyen L., Dionne D., Rozenblatt-Rosen O., Tata P.R., Mou H., Shivaraju M., Bihler H., Mense M., Tearney G.J., Rowe S.M., Engelhardt J.F., Regev A, & Rajagopal J. (2018). A revised airway epithelial hierarchy includes CFTR-expressing ionocytes. Nature, 560(7718), 319-324.
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