The fetal lung fibroblast IMR 90 cell line was purchased from ATCC (Manassas, VA). 6-well collagen I coated plates were obtained from BD Biosciences (Bedford, MA). ACE-2 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). αSMA, NCadherin, β-catenin and β-actin antibodies were purchased from Cell Signaling Technology (Danvers, MA). Oxygen delivery was confirmed in the hypoxia/hyperoxia chamber outlet using a Maxtec MaxO2+A Oxygen Analyzer - R217P62 (Salt Lake City, Utah). Protease Inhibitor Cocktail P840 was purchased from Sigma (Saint Louis, MO) and the Pierce BCA Protein Assay Kit from Thermo Fisher Scientific Pierce Biotechnology (Rockford, IL), 10X Tris/glycine/SDS buffer was obtained from BioRad (Berkeley, CA), and HRP-conjugated goat anti-rabbit secondary antibody from Santa Cruz Biotechnology (Santa Cruz, CA). All other materials were of reagent grade.
Imr 90 cell line
Manufactured by American Type Culture Collection
The IMR-90 cell line is a normal human diploid fibroblast cell line derived from fetal lung tissue. It is a widely used and well-characterized research tool for studying cellular processes and evaluating the effects of various treatments or environmental factors on cell behavior.
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Lab products found in correlation
Dmem f12, by American Type Culture Collection (2 mentions)
Eagle s mem, by American Type Culture Collection (2 mentions)
Htert rpe 1 cell line, by American Type Culture Collection (2 mentions)
Hrp conjugated goat anti rabbit secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Protease inhibitor cocktail p840, by Merck Group (1 mentions)
α sma, by Cell Signaling Technology (1 mentions)
Tris glycine sds buffer, by Bio-Rad (1 mentions)
β catenin, by Cell Signaling Technology (1 mentions)
N cadherin, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Dmem f12 medium, by Corning (1 mentions)
A549 cell line, by American Type Culture Collection (1 mentions)
F12k medium, by Boster Bio (1 mentions)
Dulbecco modified eagle medium (dmem), by Sartorius (1 mentions)
60 mm tissue culture dishes, by BD (1 mentions)
Glutamine, by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
5 protocols using imr 90 cell line
1
Fetal Lung Fibroblast Hypoxia/Hyperoxia Protocol
2
TGF-β-Induced Epithelial-Mesenchymal Transition
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The A549 cell line and IMR-90 cell line were purchased from ATCC. A549 cells were cultured in F-12K medium (Boster Biological Technology) containing 10% foetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin. IMR-90 cells were cultured in DMEM (Biological Industries) containing 10% FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin. The MLE-12 cell line was purchased from the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China). MLE-12 cells were cultured in DMEM/F-12 medium (Corning) containing 10% FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin. A549, MLE-12, and IMR-90 cells were incubated in a humidified incubator with 95% air and 5% CO2 at 37 °C. A549 cells, MLE-12 cells, and IMR-90 cells were seeded in six-well plates and cultured overnight. Cells were treated with TGF-β (10 ng/mL) for 24 h[37 (link)] and then exposed to 680C91 (20 μm) for 24 h.
3
Cell Culture and Fixation Protocol
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The IMR-90 cell line and hTERT-RPE-1 cell line were purchased from American Type Culture Collection (ATCC, cat. no. CCL-186 for IMR-90 and cat. no. CRL-4000 for hTERT-RPE-1). IMR-90 cells were cultured with Eagle’s MEM (ATCC, cat. no. 30-2003) containing 10% (vol/vol) FBS and 1× penicillin-streptomycin. The hTERT-RPE-1 cells were cultured with DMEM: F12 (ATCC, cat.no. 30-2006) medium containing 10% (vol/vol) FBS and 1× penicillin-streptomycin. For imaging, cells were plated onto 40-mm-diameter, #1.5 coverslips in Falcon 60-mm tissue culture dishes. When the cells reached a confluency of 70%, the cells were fixed with 4% (wt/vol) paraformaldehyde (PFA) in DPBS for 10 min at room temperature and washed twice with Dulbecco’s phosphate-buffered saline (DPBS). The cell lines were not authenticated or tested for mycoplasma contamination.
4
Cell Culture and Fixation Protocol
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The IMR-90 cell line and hTERT-RPE-1 cell line were purchased from American Type Culture Collection (ATCC, cat. no. CCL-186 for IMR-90 and cat. no. CRL-4000 for hTERT-RPE-1). IMR-90 cells were cultured with Eagle's MEM (ATCC, cat. no. 30-2003) containing 10% (vol/vol) FBS and 1 penicillin-streptomycin. The hTERT-RPE-1 cells were cultured with DMEM: F12 (ATCC, cat.no. 30-2006) medium containing 10% (vol/vol) FBS and 1 penicillin-streptomycin. For imaging, cells were plated onto 40-mm-diameter, #1.5 coverslips in Falcon 60-mm tissue culture dishes. When the cells reached a confluency of 70%, the cells were fixed with 4% (wt/vol) paraformaldehyde (PFA) in DPBS for 10 min at room temperature, and washed twice with Dulbecco's phosphate-buffered saline (DPBS).
5
Fibroblast and Stem Cell Culture
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The IMR90 cell line (fetal lung fibroblasts; CCL-186) was obtained from the American Type Culture Collection. The NHDF1 cell line (neonatal human dermal fibroblasts) which was originally obtained from Lonza (Allendale, NJ) was a gift of Dr Lowry (University of California at Los Angeles) [34] (link). The human fetal fibroblasts (HFF) were isolated from fetal foreskin tissues and were previously used to generate iPSCs in the lab [35] (link). IMR90 and HFF cells were maintained in fibroblast medium: DMEM supplemented with 10% fetal calf serum, glutamine and non-essential amino-acids (Life Technologies, Carlsbad, CA, USA). NHDF1 cells were maintained as described by Lowry et al [34] (link). The H1 human embryonic stem cell (H1-hESC) line was obtained from WiCell (Madison, Wisconsin, USA) and was maintained in mTeSR1 medium (Stem Cell Technologies, Vancouver, BC, Canada) on matrigel (BD Biosciences, San Jose, CA, USA) coated plates.
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