The gill cell line RTgill-W1, derived from rainbow trout Oncorhynchus mykiss [32 ], was obtained from the American Type Culture Collection (CRL-2523). This cell line was routinely cultured on 25-cm2 culture treated flasks (690170, Greiner Bio-One) with Leibovitz’s medium (L1518, Sigma) supplemented with 10% fetal bovine serum (v/v) (12003C, Sigma) and an antibiotic-antimycotic solution (A5955, Sigma). Cells were incubated at 20°C in the dark. This epithelial cell line was detached with 0.25% trypsin-0.02% EDTA solution (59428C, Sigma) for subculturing and seeding purposes [30 (link)].
L1518
Manufactured by Merck Group
The L1518 is a piece of laboratory equipment manufactured by Merck Group. It serves as a general-purpose tool for various laboratory applications. The core function of the L1518 is to provide a reliable and consistent platform for conducting experiments and analyses. No further details about the intended use or specific applications of this product are provided.
Automatically generated - may contain errors
6 protocols using l1518
1
Culturing Rainbow Trout Gill Cells
2
Culturing Rainbow Trout Gill Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell line RTgill-W1, isolated from gill filaments of rainbow trout Oncorhynchus mykiss (Bols et al., 1994) , was acquired from the American Type Culture Collection (CRL-2523, ATCC). Cells were cultured at 17°C in the dark in Leibovitz's L-15 medium (L1518 Sigma), supplemented with 10% (v/v) fetal bovine serum (FBS, 12003C, Sigma), and an antibiotic-antimycotic solution (A5955, Sigma) containing streptomycin (10 mg mL -1 ), amphotericin B (25 mg mL -1 ) and penicillin (10,000 units mL -1 ) in 25-cm 2 culture-treated flasks (3100-025, Iwaki). 0.25% trypsin-0.02% EDTA in Hank's balanced salt solution (59428C, Sigma) was used to remove cells that grew as an adherent monolayer at the bottom of the flask. Subcultures were normally established twice per week at a ratio of 1:2 with L-15 medium renewal.
3
2D Chemotaxis Assay Using H2O2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
2D chemotaxis assay was performed using µ-Slide Chemotaxis (80326, ibidi GmbH, Martinsried, Germany) as instructed. B16-F10 cells were washed with DPBS, trypsinized and centrifuged at 1000 rpm for 3 min followed by resuspension with fresh culture medium. Resuspended cells were placed into the channel between the two reservoirs. After attachment, the reservoirs were filled with CO2-independent media (L1518, Sigma-Adrich) with or without chemoattractant (50 μM H2O2). Cell motions were captured every 20 min for 9 h. Images were analyzed using “Manual tracking” and “Chemotaxis tool” plugin of ImageJ.
4
Derivation and Culture of Xenopus Epithelial Cell Line
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell line XL-177 was derived from tadpole epithelium of X. laevis41 . Cultured cells were maintained in medium containing L-15 (SIGMA L1518), sterile water and FBS (6:3:1vol/vol) and supplemented with 100 μl penicillin/streptomycin. Cells were grown at 23 °C in gelatin-coated dish that were sealed. Confluent culture of cells were split 1:3 to 1:6 following trypsin digestion.
5
PBMC Isolation from Whole Blood
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plasma and peripheral blood mononuclear cells (PBMCs) were separated immediately following manufacturer instructions (Sigma‐Aldrich, Z642843). 20 mL of whole blood was transferred from the EDTA tubes into LeucoSep-tube containing ficoll-hypaque at a ratio of 2:1. The tube was centrifuged at 800×g for 30 min at room temperature in a swinging-bucket rotor with no break. The top layer of plasma was removed, and the buffy coat interface was collected, washed twice with PBS-EDTA (10 mM), and centrifuged for 10 min at 250×g with the brake on. The pelleted cells were suspended in red blood cell lysis buffer (1 mM KHCO3, 0.15 M NH4Cl, 0.1 mM EDTA, HCl pH 7.2 to 7.4) at room temperature for 5 min. The cells were washed again with PBS-EDTA, centrifuged at 250×g for 10 min at 4 °C and resuspended in appropriate medium (Leibovitz medium, Sigma-Aldrich, L1518) for further assay (ELISpot). The plasma was centrifuged at 250×g for 5 min at 4 °C and transferred to a new 15 mL tube to remove cells and debris. Both the PBMCs and plasma were transferred to 2 mL cryotubes for further assay (ELISA) and storage at − 80 °C.
6
Culturing Breast Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human breast cancer cell lines (MDA-MB-231, MDA-MB-468, SKBR3, BT474, MCF7) were cultured in L15 (L1518; Sigma Aldrich), Dulbecco’s Modified Eagle Medium (DMEM) (D6429; Sigma Aldrich) or 5A (M8403; Sigma Aldrich) culture medium supplemented with 10% fetal bovine serum (Hyclone). The human mammary epithelial cell line MCF10A was cultured in Mammary Epithelial Cell Medium (CM-0525, Procell, China).
Cell lines were authenticated by short tandem repeat (STR) profiling and were tested negative for Mycoplasma.
Cell lines were authenticated by short tandem repeat (STR) profiling and were tested negative for Mycoplasma.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!