Canine aortic arches (n = 30) were harvested under sterile conditions from euthanized dogs (foxhounds) of other ongoing experimental studies. After the separation of adhesive tissue, all samples were examined macroscopically to exclude any pathology and stored in Medium 199 with Earle's salts (PAA Laboratories GmBH, Pasching, Austria) containing 10% dimethyl sulfoxide (DMSO) at -80°C until further use. Aortic arches were decellularized through continuous shaking in 1% sodium dodecyl sulfate (SDS) and 0.05% sodium azide (NaN3) in phosphate buffered saline (PBS) (PAA Laboratories) at room temperature for 48 h. The solution was exchanged every 6 h. At the end of the decellularization protocol, the aortic arches were washed with PBS for 12 h to remove residual detergents and cell debris, then stored in 1% penicillin-streptomycin (PAA Laboratories, Cölbe, Germany) augmented Earle's Medium 199 (PAA Laboratories) until implantation/other measurements (Figure S1 , left side).
Earle s salts
Manufactured by GE Healthcare
Sourced in Austria, United Kingdom
Earle's salts is a cell culture medium used to support the growth and maintenance of various cell types in vitro. It provides a balanced salt solution with essential inorganic ions, such as calcium, magnesium, and potassium, to maintain the osmotic pressure and pH required for cell viability.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by GE Healthcare (3 mentions)
Mccoy s 5a medium, by Biowest (2 mentions)
60 mm culture plates, by Corning (2 mentions)
Hct 116 cells, by Leibniz Institute DSMZ (2 mentions)
Ultra low attachment plate, by Corning (2 mentions)
6 well culture plate, by Corning (2 mentions)
Phosphate buffered saline (pbs), by GE Healthcare (1 mentions)
Penicillin streptomycin, by GE Healthcare (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Caco 2, by GE Healthcare (1 mentions)
Caco 2, by American Type Culture Collection (1 mentions)
Hrt18, by American Type Culture Collection (1 mentions)
Dmem high glucose medium, by Capricorn (1 mentions)
Dld 1 cells, by American Type Culture Collection (1 mentions)
L glutamine, by GE Healthcare (1 mentions)
Penicillin, by GE Healthcare (1 mentions)
Streptomycin, by GE Healthcare (1 mentions)
Mycoalert mycoplasma detection kit, by Lonza (1 mentions)
5 protocols using earle s salts
1
Decellularized Canine Aortic Arches
2
Culturing Colorectal Cancer Cell Lines
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Caco-2 cells (ECACC, Salisbury, UK) were grown in MEM with Earle’s salts (PAA Laboratories GmbH, Pasching, Austria) containing 15% fetal bovine serum. HCT-116 cells (DSMZ, Braunschweig, Germany) were grown in McCoy’s 5A medium (Biowest, Nuaillé, France) containing 10% fetal bovine serum (PAA Laboratories). HCT-116 cells harbour an activating mutation in KRAS (G13D), while Caco-2 cells are KRAS wildtype. Culture media were supplemented with 2 mM glutamine, 1% non-essential amino acids, penicillin (100 U/ml), streptomycin (100 μg/ml) and amphotericin B (2.5 μg/ml). Cells were maintained in a humidified atmosphere at 37°C and 5% CO2. After cultures became 80% confluent (usually after 3 days), cells were trypsinized, centrifuged, and suspended in fresh medium. All cells used for experiments displayed > 95% viability and were seeded in 96-well plates, 6-well culture plates, 60-mm culture plates or ultra low-attachment plates (Corning Inc, Lowell, MA, USA). All experiments were carried out in duplicate and repeated at least three times.
3
Caco-2 and HRT-18 Cell Culture
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The CRC cell lines Caco-2 (KRAS wild-type) and HRT-18 (KRAS-mutated) were purchased from American Type Culture Collection. Caco-2 was cultured in MEM with Earle’s salts (PAA Laboratories, Pasching, Austria) supplemented with 2 mmol/L of l -glutamine, 1.5 g/L of sodium bicarbonate, 0.1 mmol/L of nonessential amino acids, 50 units/mL of penicillin, 50 µg/mL of streptomycin, and 10% FBS (PAA Laboratories). HRT-18 cells were maintained in RPMI 1640 (GIBCO, Vienna, Austria) containing 2 mmol/L of l -glutamine, 50 units/mL of penicillin, 50 µg/mL of streptomycin, and 10% FBS. After obtaining a confluence of approximately 80%, total RNA was isolated following a standard TRIzol protocol and RNA was stored at −70 °C until further procedures.
4
Culturing of Colorectal Cancer Cell Lines
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HT-29 and HCT-116 cells (DSMZ, Braunschweig, Germany) were grown in McCoy's 5A medium (Biowest, Nuaillé, France) containing 10% fetal bovine serum (PAA Laboratories, Pasching, Austria). Caco-2 cells (ECACC, Salisbury, UK) were grown in MEM with Earle's salts (PAA Laboratories GmbH) containing 15% fetal bovine serum. DLD-1 cells (ATCC, US ) were grown in D-MEM high glucose medium (Capricorn scientific, GmbH), containing 15% fetal bovine serum. Culture media were supplemented with 2 mM glutamine, 1% non-essential amino acids, penicillin (100 U/ml), streptomycin (100 μg/ml) and amphotericin B (2.5 μg/ml) and cells were cultured in a humidified atmosphere at 37°C and 5% CO2. in 96-well plates, 6-well culture plates, 60-mm culture plates or ultra low-attachment plates (Corning Inc, Lowell, MA, USA). All experiments were carried out in duplicate and repeated at least three times.
5
Culturing F508del CFTR Mutant Cell Lines
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The CFBE41o-human bronchial epithelial (homozygous for the F508del mutation) [29] and ΣCFTE29ohuman tracheal epithelial (homozygous for the F508del mutation) [30] cell lines carrying the F508del CFTR mutation (so called CF cells) and the 16HBE14onormal human bronchial epithelial cell line [31] (generous gifts from Dieter Gruenert, San Francisco, CA, U.S.A.) were cultured at 37°C in a 5% CO 2 -humidified incubator in 20 ml MEM with Earle's Salts (PAA Laboratories), 0.4% Penicillin (40 Units/ml)/Streptomycin (40 µg/ml) (PAA Laboratories), 2mM L-Glutamine (PAA Laboratories), supplemented with 10% heat-inactivated fetal bovine serum (PAA Laboratories). Tissue culture plastic wares (75 cm 2 ) were coated for 20-30 min at 37°C with MEM with Earle's Salts containing fibronectin (0.01 mg/ml), collagen (0.03 mg/ml) and bovine serum albumin (BSA) (0.1 mg/ml). The culture medium was changed every 2 days. The absence of mycoplasma in cell cultures was determined by using MycoAlert® Mycoplasma Detection Kit (Lonza, Levallois Perret, France).
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