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Laminin 1

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Laminin 1 is a glycoprotein that is a major component of the basement membrane. It plays a crucial role in cell attachment, differentiation, migration, and survival.

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Lab products found in correlation

Cell culture microplate, by Agilent Technologies (2 mentions) Oligomycin, by Merck Group (2 mentions) Seahorse xf96 flux analyzer, by Agilent Technologies (2 mentions) Carbonyl cyanide 4 trifluoromethoxy phenylhydrazone fccp, by Merck Group (2 mentions) Xf assay medium, by Agilent Technologies (2 mentions) Antimycin a, by Merck Group (2 mentions) Rotenone, by Merck Group (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Lsm 880, by Zeiss (2 mentions) Parafilm m, by Amcor (1 mentions) Hcs cellmask deep red stain, by Thermo Fisher Scientific (1 mentions) Sorvall legend rt, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions) Triton x 100, by Merck Group (1 mentions) Phosphate buffered saline (pbs), by Corning (1 mentions)

4 protocols using laminin 1

1

Seahorse Bioenergetic Analysis of COPD Airway Cells

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Extracellular acidification rates and oxygen consumption rates were determined by the Seahorse XF 96 flux analyzer (Seahorse Bioscience). Primary airway epithelial cells were isolated as described above and plated onto cell culture microplates (Seahorse Bioscience) coated with 50 ng/μl Laminin 1 (3400-010-01, Trevigen) for 4 days (media change on Day 2). On the day of the experiment cells were treated with CSE for 4 hours. Cells were incubated in XF assay medium (Seahorse bioscience), supplemented with 5 mM glucose, 4 mM glutamine and 1 mM pyruvate for one hour prior to the measurement. After the recording of the basal rates of ECAR and OCR, final concentrations of 1 μM oligomycin, 2 μM carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP) and 0.5–0.5 μM rotenone and antimycin A were added (Sigma) through the instruments injection ports in order to obtain proton leak, maximal respiratory capacity and non-mitochondrial respiration respectively. Rates were normalized by DNA with Hoechst 33342 against standards with known concentrations of DNA.
Cloonan S.M., Glass K., Laucho-Contreras M.E., Bhashyam A.R., Cervo M., Pabón M.A., Konrad C., Polverino F., Siempos I.I., Perez E., Mizumura K., Ghosh M.C., Parameswaran H., Williams N.C., Rooney K.T., Chen Z.H., Goldklang M.P., Yuan G.C., Moore S.C., Demeo D.L., Rouault T.A., D’Armiento J.M., Schon E.A., Manfredi G., Quackenbush J., Mahmood A., Silverman E.K., Owen C.A, & Choi A.M. (2016). Mitochondrial iron chelation ameliorates cigarette-smoke induced bronchitis and emphysema in mice. Nature medicine, 22(2), 163-174.
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2

Seahorse Bioenergetic Analysis of COPD Airway Cells

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Extracellular acidification rates and oxygen consumption rates were determined by the Seahorse XF 96 flux analyzer (Seahorse Bioscience). Primary airway epithelial cells were isolated as described above and plated onto cell culture microplates (Seahorse Bioscience) coated with 50 ng/μl Laminin 1 (3400-010-01, Trevigen) for 4 days (media change on Day 2). On the day of the experiment cells were treated with CSE for 4 hours. Cells were incubated in XF assay medium (Seahorse bioscience), supplemented with 5 mM glucose, 4 mM glutamine and 1 mM pyruvate for one hour prior to the measurement. After the recording of the basal rates of ECAR and OCR, final concentrations of 1 μM oligomycin, 2 μM carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP) and 0.5–0.5 μM rotenone and antimycin A were added (Sigma) through the instruments injection ports in order to obtain proton leak, maximal respiratory capacity and non-mitochondrial respiration respectively. Rates were normalized by DNA with Hoechst 33342 against standards with known concentrations of DNA.
Cloonan S.M., Glass K., Laucho-Contreras M.E., Bhashyam A.R., Cervo M., Pabón M.A., Konrad C., Polverino F., Siempos I.I., Perez E., Mizumura K., Ghosh M.C., Parameswaran H., Williams N.C., Rooney K.T., Chen Z.H., Goldklang M.P., Yuan G.C., Moore S.C., Demeo D.L., Rouault T.A., D’Armiento J.M., Schon E.A., Manfredi G., Quackenbush J., Mahmood A., Silverman E.K., Owen C.A, & Choi A.M. (2016). Mitochondrial iron chelation ameliorates cigarette-smoke induced bronchitis and emphysema in mice. Nature medicine, 22(2), 163-174.
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3

Lacrimal Gland Progenitor Cell Culture

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We used a spheroid culture method modified from one for neural stem cells, and then a floating organoid culture method modified from one used for thymic epithelial cells and lacrimal gland epithelium as we reported previously.6 Briefly, after isolation, lacrimal gland progenitor cells were resuspended in culture medium as 4 × 106 cells/mL, and 50 μL of cell suspension was seeded in each micro-mold (Microtissues, Inc., Providence, RI). The formed spheroids were cultured in low attachment flasks for one more day, and then seeded into a 15 μL drop of laminin I (6 mg/mL; Trevigen, Inc., Gaithersburg, MD) sitting on a polycarbonate filter floating in serum-free medium for 2 weeks.
Lin H., Liu Y, & Yiu S. (2019). Three Dimensional Culture of Potential Epithelial Progenitor Cells in Human Lacrimal Gland. Translational Vision Science & Technology, 8(4), 32.
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4

3D Printed Microfluidic Device for Cell Culture

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Corning Costar 7007 ultra-low attachment round bottom 96-well plates and phosphate buffered saline (PBS) were obtained from Corning (Corning, NY). Dulbecco’s Modified Eagle Medium (DMEM), penicillin-streptomycin, and HCS CellMask Deep Red stain were acquired from Thermo Fisher Scientific (Waltham, MA). Laminin-I was purchased from Trevigen (Gaithersburg, MD). For fixation, 8% paraformaldehyde (PFA) solution was obtained from Electron Microscopy Sciences (Hatfield, PA). Parafilm “M” was obtained from Bemis (Oshkosh, WI). Fetal bovine serum (FBS), Agar powder, and Triton X-100 were purchased from Sigma-Aldrich (St. Louis, MO). The High-Density Polyethylene (HDPE) inserts were acquired from Hero Glue (Las Vegas, NV). The neoprene gasket, absorbent mat, and nylon mesh for the containment base were obtained from McMaster-Carr (Elmhurst, IL). All other device components were fabricated with a Stratasys (Rehovot, Israel) Objet 30 Polyjet 3D printer using VeroWhitePlus acrylic photopolymer and support material. Centrifugation was conducted using a Sorvall Legend RT obtained from Thermo Fisher Scientific (Waltham, MA). For confocal microscopy, a Zeiss (Oberkochen, Germany) LSM 880 was used.
Weisler W., Miller S., Jernigan S., Buckner G, & Bryant M. (2020). Design and testing of a centrifugal fluidic device for populating microarrays of spheroid cancer cell cultures. Journal of Biological Engineering, 14, 7.
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