Caco-2 cells were seeded into the wells of a 48-well plate, coated with 0.1% bovine collagen (Sigma-Aldrich) for 24 h, 3 weeks before the experiment with complete DMEM/F12 (1:1) medium renewal every 2–3 days. Monocytes, moDCs, CD4+ T-helper and CD8+ T cytotoxic cells, Mφ1, and Regulatory CD4+ FoxP3+ T cells were seeded into the wells of 48-well plates in the complete RPMI-1640 medium supplemented with 10% Human AB serum (HABS, Capricorn Scientific, Ebsdorfergrund, Germany) and 1XAAS 24 h prior to the experiment; medium in Caco-2-containing wells was also replaced with the same medium. After 24 h, half of the medium in each well was replaced by fresh complete RPMI-1640 medium with 10% HABS, supplemented with 4 µM of Lc-def for the sample wells or fresh medium alone for the control wells. Cell cultures were kept in CO2-incubator (5% CO2, 37 °C) for another 24 h. Culture supernatants were collected 24 h later and stored at −70 °C degrees less than one month prior to the analytes assessment.
Bovine collagen
Manufactured by Merck Group
Sourced in Germany, United States
Bovine collagen is a type of lab equipment used in various research and testing applications. It is derived from the connective tissue of cattle. Bovine collagen is a structural protein that provides support and structure to tissues.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Pustulan, by Merck Group (1 mentions)
Laminarin, by Merck Group (1 mentions)
Chitin, by Merck Group (1 mentions)
Laminarin, by Thermo Fisher Scientific (1 mentions)
β 1 3 glucan, by Thermo Fisher Scientific (1 mentions)
Tryple express, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Glutamine, by Thermo Fisher Scientific (1 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions)
Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (1 mentions)
Sylgard 184 silicone elastomer kit, by Dow (1 mentions)
Epinephrine, by Merck Group (1 mentions)
6 protocols using bovine collagen
1
Immune cell co-culture with Caco-2 cells
2
Hyaluronan-Gelatin Scaffolds for Cell Carriers
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Sponge scaffolds were formulated as a cell carrier for the study and manufactured from 70% derivatized hyaluronan-ester and 30% gelatin, as previously described [25 (link), 26 (link)]. The hyaluronan component was obtained from the commercially available product, Jaloskin (Fidia Advanced Biopolymers, Abano Terme, Italy), which is manufactured from sodium hyaluronate and highly esterified with benzyl alcohol on the free carboxyl groups of glucoronic acid within the polymer. The gelatin component was hydrolyzed bovine collagen (Sigma, Taufkirchen, Germany). Porous scaffolds were manufactured by solvent casting via a particulate leaching technique, using NaCl with controlled grain size as the porogen. The primary pore size was 250–350 μm and secondary pore size was 50–100 μm. Scaffolds had a diameter of 2.2 mm and a height of 3 mm.
3
Binding Capacity of C. glabrata to Fungal Cell Wall
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Binding capacity of C. glabrata cells to fungal cell wall components and extracellular matrix (ECM) collagen was also evaluated by flow cytometry following a similar procedure as described above for 4 h adhesion to polystyrene, using the same amount of cells. However, before incubation with C. glabrata, the microtiter plates were coated with chitin (Sigma; 250 μg/mL in 1% acetic acid), laminarin (β-1,3-glucan, Thermo Fisher; 500 μg/mL in milli-Q water), pustulan (β-1,6-glucan, Calbiochem; 500 μg/mL in 50 mM potassium acetate buffer) or bovine collagen (Sigma; 250 μg/mL in 0.2 M bicarbonate buffer, pH 9.6) by adding 500 μL of the respective solutions, and allowing for passive adsorption (1 h at 30°C followed by overnight incubation at 4°C), in the case of chitin and collagen, or evaporation (overnight at 37°C), in the case of laminarin and pustulan.
4
Muscle Biopsy Explant Culture Protocol
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Muscle biopsies were cultured in multi-well dishes coated with 0.1% bovine collagen (Sigma-Aldrich for all of the compounds, as declared elsewhere) at 37°C in a 5% CO2 and 5% O2 atmosphere according to the explant technique (Cusella-De Angelis et al., 1994 (link); Messina et al., 2004 (link); Morosetti et al., 2006 (link); Tonlorenzi et al., 2007 (link); Díaz-Manera et al., 2012 (link); Quattrocelli et al., 2012 (link)), as direct enzymatic digestion of the microbiopsy resulted in too low yields for stem cell-derived progenitor isolation, culture and further experiments. Growth medium containing IMDM (Thermo Fisher Scientific) was used with 20% fetal bovine serum (FBS; Thermo Fisher Scientific), 1 mg/mL non-essential aminoacids (Thermo Fisher Scientific), 1 mg/mL sodium-pyruvate (Thermo Fisher Scientific), 1 U/mL penicillin/streptomycin (P/S; Thermo Fisher Scientific), 1% chicken embryo extract (CEE; Bio Connect), 2 mM Glutamine (Thermo Fisher Scientific) and 100 nM β-mercaptoethanol (Thermo Fisher Scientific). When 70–80% of confluence was reached, cells were passaged, using TrypLETM Express (Thermo Fisher Scientific). After cell sorting and amplification, cells were seeded at different densities to test their differentiation potential using specific differentiation media to induce different cell fate programs, as specified in “In vitro differentiation assays.”
5
Fabrication of Anisotropic hDF-ECM Sheets
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A silicon wafer with aligned grooves (width: 5 μm, Pitch: 8 μm, height: 270 nm) was prepared with a soft lithography technique at the microfabrication shared facility (MFF) at Michigan Technological University. Polydimethylsiloxane (PDMS) molds were cased from the silicon wafer using Sylgard 184 Silicone Elastomer Kit (Dow Corning, Midland, MI) by mixing base and cross-linker at 10:1 ratio followed by heating at 65 °C for 4 h. Flat PDMS was cast from a flat plastic petri dish. Micro-patterned and flat PDMS were coated with polydopamine and collagen-I prior to cell culture as described in the previous publication [68 (link)]. Briefly, PDMS were immersed in 0.01% W/V 3-hydroxytyramine hydrochloride (Dopamine-HCl) (ACROS Organics, Fisher scientific) for 24 h followed by ethylene oxide sterilization. Polydopamine coated PDMS were immersed in bovine collagen (20 μg/mL) (Sigma Aldrich, St. Louis, MO) for 2 h before cell seeding [68 (link)]. Neonatal human dermal fibroblasts (ATCC, Manassas, VA) (passage 3–5) were cultured on micropatterned and flat PDMS for 6 weeks to develop anisotropic or isotropic hDF sheets, which were then decellularized to fabricate anisotropic or isotropic hDF-ECM as described previously [14 (link),69 ]. Briefly, hDF sheets were decellularized using 1 M NaCl, 0.5% Sodium dodecyl sulfate (SDS), 10 mM Tris and 5 mM ethylenediaminetetraacetate (EDTA) [14 (link)].
6
Preparation and Evaluation of Boswellic Acid
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Boswellic acid was purchased from Advance Physician Formulas Inc. (California, USA) in tablet form; it was dissolved in distilled water. Acetyl salicylic acid was kindly provided by Medical Union Pharmaceuticals (MUP, Ismailia, Egypt). Bovine collagen and epinephrine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cholesterol was purchased from GFS chemicals (Texas, USA) and bile salts were purchased from SAS Chemicals Co. (Mumbai, India). All the used solvents were of analytical grade and were supplied by Al-Nasr Company for Chemical Industries (Cairo, Egypt).
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