For DC differentiation, 0.5 to 1.0 × 106 monocytes/mL were cultured for five days in RPMI medium supplemented with 10% fetal calf serum (PAN Biotech), IL-4 (144 U/mL), and granulocyte macrophage colony-stimulating factor (GM-CSF, 225 U/mL; both from PeproTech, Hamburg, Germany). iDCs were then stimulated with 100 ng/mL LPS (from Salmonella abortus equi S-form, Enzo Life Sciences, Lörrach, Germany), 25-hydroxyvitamin D3 (Sigma-Aldrich) (25 nM to 100 nM) and or ATG (Fresenius, Bad Homburg, Germany) (now named Grafalon®, distributed by Neovii Biotech, Gräfelfing, Germany) (100 µg/mL), Cyclosporine A (Sandimmun, Novartis), Dexamethasone (Jenapharm, mibe GmbH), IgG isotype control (polyclonal, rabbit, Molecular Innovations, Novi, MI, USA) (100 µg/mL) for 48 hours.
Cyclosporine a
Manufactured by Novartis
Sourced in Switzerland, United States, Germany, Japan
Cyclosporine A is a laboratory reagent used in biochemistry and molecular biology research. It is a cyclic peptide that inhibits the activity of calcineurin, a key enzyme involved in the activation of T cells. Cyclosporine A is commonly used as a tool to study signal transduction pathways and immune system regulation.
Automatically generated - may contain errors
Lab products found in correlation
Pinacidil, by Merck Group (3 mentions)
Sandimmune, by Novartis (3 mentions)
Interleukin 4 (il 4), by R&D Systems (2 mentions)
Igf 1, by Thermo Fisher Scientific (2 mentions)
Cryostor, by Merck Group (2 mentions)
Bcl xl bh4, by Merck Group (2 mentions)
Cyclosporine, by Novartis (2 mentions)
Interleukin 4 (il 4), by PAN Biotech (1 mentions)
Igg isotype control, by Innovative Research (1 mentions)
Gm csf, by Thermo Fisher Scientific (1 mentions)
25 hydroxyvitamin d3, by Merck Group (1 mentions)
Lps from salmonella abortus equi s form, by Enzo Life Sciences (1 mentions)
Xylazin, by Bayer (1 mentions)
Tissue tek, by Sakura Finetek (1 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Z vad fmk, by Promega (1 mentions)
Mitomycin c mmc, by Fujifilm (1 mentions)
Tacrolimus, by Novartis (1 mentions)
Atopica, by Novartis (1 mentions)
48 protocols using cyclosporine a
1
Differentiation and Maturation of Dendritic Cells
2
Cyclosporine A Dosing in Rat Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
3
Rat Orthotopic Kidney Transplantation Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 15 male Fischer (F344) weighing 150–220 g, aged 8–12-week-old were used for this procedure. A surgical ASX-2 microscope was purchased from Shanghai Anxin Optical Instrument Manufacture Co., Ltd. (Shanghai, China). Orthotopic kidney transplantation was performed as previously described (9 (link)). Surgery was performed under general anesthesia with 3% sodium pentobarbital (30 mg/kg) administered via intraperitoneal (i.p.) injection. The left kidney from the recipient Lew rat was removed and the left renal vessels were clamped. The left donor kidney from a F344 rat was then removed, cooled and positioned orthotopically in the recipient Lew rat. The renal arteries, veins and ureters of the donor and recipient were then anastomosed end-to-end with 10-0 prolene (Ningbo Lingqiao Biological Technology, Ningbo, China). No ureteral stents were used. A low dose of cyclosporine A (1.5 mg/kg/day; Novartis International AG, Basel, Switzerland) was administered for 10 days following transplantation to suppress acute rejection. To prevent infection, recipients received ceftriaxone (Rocephin; 20 mg/kg/day) via i.p. injection for 3 days. The right native kidney of each recipient was removed 10 days following surgery. Rats exhibiting overt signs of unsuccessful surgeries were excluded from the current study. Subsequently there were 10 rats per group following exclusion.
4
Collagen Scaffold Implantation in Mouse Hearts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Animals underwent a left lateral thoracotomy after intraperitoneal ketamine (100 mg/kg; Merial, Gerland, France)-xylazin (10 mg/kg; Bayer, Puteaux, France) anaesthesia and tracheal ventilation. Analgesia was performed for 2 days after surgery with a 2 mg/kg intraperitoneal injection of profenid® (Merial). Collagen scaffolds (surface approximately 200 mm²) were sutured on the ventricles of 24 healthy C57BL6 and 26 αMHC-MerCreMer:Sf/Sf mice. Six additional αMHC-MerCreMer:Sf/Sf mice (Sham) underwent the same procedure without scaffold implantation and served as sham-operated controls. All αMHC-MerCreMer:Sf/Sf mice were daily treated with cyclosporine A (10 mg/kg/day, intraperitoneal; Novartis Pharma, Rueil-Malmaison, France) until the echocardiographic assessment of heart function at 14 days after the implantation. C57BL6 mice were sacrificed 7 days after treatment to assess the biocompatibility of collagen scaffold. All mice were sacrificed by cervical dislocation and then hearts were removed, the left ventricle apexes were dissected and flash-frozen in liquid nitrogen for subsequent RNA isolation. The remaining hearts were embedded in Tissue-Tek (Sakura Finetek, Villeneuve d'Ascq, France) and frozen in liquid nitrogencooled isopentane until they were sliced into 10 µm thick cryosections for histology and immunostaining (see Supplemental Methods).
5
Immortalization of B Cells from Humanized Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Splenocytes obtained from humanized mice were dissociated into single cell suspensions and used immediately. Infection was performed with the EBV virus strain B95.8 at a multiplicity of infection (MOI) of 0.1. 5 × 106 splenocytes were incubated for 3 h at 37 °C and 5% CO2 with EBV in RPMI 1640 media (Gibco, Thermo Fisher, Waltham, MA, USA), supplemented with FBS (described above), Cyclosporine A (CsA, 1µg/mL, Novartis, Basel, Switzerland) and IL-4 (2 ng/mL R&D Systems, Minneapolis, MN) and 1% penicillin/streptomycin (Gibco, Thermo Fisher, Waltham, MA, USA). Irradiated feeder cells prepared on the day prior to the EBV infection were used to harness immortalization. LL8 and LEXL5 cells were irradiated at 180 Gy in a 137Cs device. 1 × 106 irradiated LL8 cells were seeded per well on 96-well cell culture plates. On the next day, 2.5 × 104 infected cells were added per well. Then, 2.5 × 104 irradiated LEXL5 cells were added per well of the cell mixture. Half of the media was replaced with newly prepared media weekly. CsA was added during the first 4 weeks of culture, to deplete the T cells from the culture. IL-4 was kept during the whole culture to improve the B cell viability and expansion.
6
Cryopreservation and Thawing of Cardiomyocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cardiomyocytes used for in vivo studies were cryopreserved on Day 20–22 of differentiation and thawed immediately prior to cell injection, following our previously described protocol (Gerbin et al., 2015 (link); Laflamme et al., 2007 (link)). One day prior to cryopreservation, cells were heat-shocked for 30 minutes at 42°C, treated with 10 μM Y-27632 for 1 hour, and dispersed with 0.25% trypsin in EDTA. Cells were spun down, resuspended in CryoStor (Sigma C2874) at 1×107 cells/mL, and frozen in cryovials. To thaw cryopreserved cells, cryovials were thawed briefly at 37°C, followed by addition of RPMI-B27 + 200 U/mL DNase. Cells were washed and resuspended in an RPMI-based pro-survival cocktail containing 50% (vol/vol) growth factor-reduced Matrigel, 100 μM ZVAD (benzylox-ycarbonyl-Val-Ala-Asp(O-methyl)-fluoro-methyl ketone, Millipore 627610), 50 nM Bcl-XL BH4 (cell-permeant TAT peptide, Millipore 197217), 200 nM cyclosporine A (Novartis), 100 ng/mL IGF-1 (Peprotech 100–11), and 50 μM pinacidil (Sigma P154).
7
Cryopreservation and Thawing of Cardiomyocytes
8
Evaluation of Cytotoxic Agents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Epirubicin (EPI) and mitomycin C (MMC) were purchased from Wako Pure Chemical Industries (Tokyo, Japan). DPQ was purchased from Abcam (Cambridge, MA, USA). Cyclosporine A (CsA) was kindly provided by Novartis Pharma K.K. (Tokyo, Japan). Z-VAD-FMK was purchased from Promega (Madison, WI, USA).
9
Avocado and Cyclosporine-A Sourcing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Avocados used in this study were obtained from a local market. Cyclosporine-A (Sandimun Neoral®) was purchased in form of capsules (25 mg/capsule) from Novartis Pharma Co., Plantation, FL, USA.
10
Calcineurin Inhibitor Effects on Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cyclosporine A and tacrolimus were purchased from Novartis Pharma Japan and Astellas Pharma Japan, respectively. For the proliferation assay, cells were cultured in medium supplemented with 0.1 μm Cyclosporine A or 0.5 μm tacrolimus until the end of the IncuCyte proliferation assay. To confirm the suppression of calcineurin activity under the treatment with Cyclosporine A and tacrolimus, cell lysates were extracted 96 h after treatment with CNIs and calcineurin activity was compared between extracted cell lines with or without the treatment of CNIs.
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!