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Iscove modified dulbecco media (imdm)

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IMDM (Iscove's Modified Dulbecco's Medium) is a cell culture medium formulated for the growth and maintenance of a variety of mammalian cell types. It is a complex and nutrient-rich medium that provides the necessary components for cell proliferation and survival.

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Lab products found in correlation

L glutamine, by Euroclone (4 mentions) Penicillin, by Euroclone (3 mentions) Interleukin 6 (il 6), by Miltenyi Biotec (3 mentions) α mem, by Thermo Fisher Scientific (3 mentions) Streptomycin, by Euroclone (2 mentions) Ccd 1064sk, by American Type Culture Collection (2 mentions) Penicillin streptomycin, by Euroclone (2 mentions) Dulbecco modified eagle medium (dmem), by Euroclone (2 mentions) Fetal bovine serum (fbs), by Euroclone (2 mentions) Mycoalert mycoplasma detection kit, by Lonza (2 mentions) Flt3l, by Miltenyi Biotec (2 mentions) Interleukin 3 (il 3), by Miltenyi Biotec (2 mentions) Iscove modified dulbecco media (imdm), by STEMCELL (2 mentions) Human serum, by Lonza (2 mentions) Heparin, by Merck Group (2 mentions) L glutamine, by Lonza (2 mentions) Fetal bovine serum (fbs), by Carlo Erba (2 mentions) Rd es, by American Type Culture Collection (2 mentions) Gentamycin, by Euroclone (1 mentions) β mercaptoethanol, by Euroclone (1 mentions)

17 protocols using iscove modified dulbecco media (imdm)

1

Isolation and Culture of Murine Dendritic Cells

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D1 cells were maintained in vitro in Iscove’s modified Dulbecco’s medium (IMDM, Euroclone) supplemented with 10% heat-inactivated fetal bovine serum (Gibco, origin: Australia), 100 IU/ml penicillin, 100 μg/ml streptomycin, 2 mM L-glutamine (all from Euroclone) and 50 μM β-mercaptoethanol (Sigma) plus 30% R1 medium (supernatant from NIH3T3 fibroblasts transfected with GM-CSF). Cells were incubated at 37 °C under 5% CO2.
Bone marrow-derived Dendritic Cells (BMDC) from C57BL/6 wild-type (WT) and MyD88 knock-out (KO) mice were cultured in IMDM (Euroclone, Milan, Italy) supplemented with 10% heat-inactivated FBS, 2 mM L-glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin (all from Euroclone), 50 μM β-mercaptoethnol (Sigma), (IMDM complete medium), and 20% supernatant of GM-CSF transduced B16 tumor cells (20 ng/ml GM-CSF). Isolated cells were seeded at 6 × 106 cells/petri dish with 10 ml of grow medium (IMDM complete, 20% B16). Cells were incubated at 37 °C under 7% CO2. At day 3, 10 ml of fresh medium were added; at day 6, 10 ml of medium were changed. After 7–10 cultivation days, cells were analyzed by Flow cytometry (FACS) for CD11c expression and used in assays when 75–85% of the cells were CD11c-positive.
Bone marrow-derived macrophages (BMDM) from C57BL/6 wild-type (WT) were generated, as previously described, by culturing cells in medium containing M-CSF37 (link).
Fumagalli S., Torri A., Papagna A., Citterio S., Mainoldi F, & Foti M. (2016). IL-22 is rapidly induced by Pathogen Recognition Receptors Stimulation in Bone-Marrow-derived Dendritic Cells in the Absence of IL-23. Scientific Reports, 6, 33900.
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2

Culturing Human Dermal Fibroblasts for Research

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Human dermal fibroblasts CCD-1064Sk (ATCC, USA) were cultured in Iscove Modified Dulbecco Media (IMDM) (Euroclone, France). The cells were supplemented with l-glutamine (1.0%, 2.0 mM), FBS (10.0% v/v), penicillin (100 U mL−1), streptomycin (100 μg mL−1), and gentamycin (1.0 mL) (Euroclone, France) at 5% CO2 and 99% relative humidity at 37 °C. The cells were stained after confluency with trypan blue dye (0.04%) and counted by a hemocytometer.
Hamad K.M., Mahmoud N.N., Al-Dabash S., Al-Samad L.A., Abdallah M, & Al-Bakri A.G. (2020). Fluconazole conjugated-gold nanorods as an antifungal nanomedicine with low cytotoxicity against human dermal fibroblasts. RSC Advances, 10(43), 25889-25897.
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3

LY27805 Cell Line Characterization

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The LY27805 cell line was kindly provided by Bruno Amati’s group [7 (link)] at the European Institute of Oncology–Italian Foundation for Cancer Research (FIRC) Institute of Molecular Oncology (IEO–IFOM, Milan, Italy) campus, expanded and stored according to their instructions. Specifically, LY27805 cells were plated at 2 × 105 cells/mL in B cell medium: 50:50 mixture of DMEM and IMDM (Euroclone, Pero (MI), Italy), 10% FBS (Euroclone, Pero (MI), Italy), 2 mM L-glutamine (Euroclone, Pero (MI), Italy), 1% penicillin/streptomycin (Euroclone, Pero (MI), Italy), 50 μM β-mercaptoethanol (Euroclone, Pero (MI), Italy), and non-essential amino acid (NEEA) (Euroclone, Pero (MI), Italy). Phoenix-Ampho cells were cultured with DMEM (Euclone, Pero (MI), Italy), 10%FBS (Euclone, Pero (MI), Italy), 2 mM L-glutamine (Euclone, Pero (MI), Italy), 1% penicillin/streptomycin (Euclone, Pero (MI), Italy).
Cells were tested and authenticated by the StemElite ID System (Promega, Madison, WI, USA). Cells were tested every six months for Mycoplasma by means of the ATCC Universal Mycoplasma Detection Kit 30-1012, cultured for no more than two weeks, and used for no longer than 15 passages.
Orecchioni S., Falvo P., Talarico G., Mitola G., Bravetti G., Mancuso P., Nicoli P, & Bertolini F. (2023). Vinorelbine and Intermittent Cyclophosphamide Sensitize an Aggressive Myc-Driven B-Cell Lymphoma to Anti-PD-1 by an Immunological Memory Effective against Tumor Re-Challenge. Journal of Clinical Medicine, 12(7), 2535.
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4

Culturing U-2OS Osteosarcoma Cells

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U-2OS human osteosarcoma cells were purchased from the American Type Culture Collection (ATCC) and grown in Iscove's modified Dulbecco's medium (IMDM, EuroClone, Milan, Italy) supplemented with 10% fetal bovine serum (Lonza, Basel, Switzerland), 2 mM L-glutamine and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin).
De Luca A., Mei G., Rosato N., Nicolai E., Federici L., Palumbo C., Pastore A., Serra M, & Caccuri A.M. (2014). The fine-tuning of TRAF2–GSTP1-1 interaction: effect of ligand binding and in situ detection of the complex. Cell Death & Disease, 5(1), e1015-.
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5

Generation of Bone Marrow-Derived Dendritic Cells

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BMDCs were generated from bone marrow precursors of C57BL/6 WT, flushed from femurs, in Iscove’s modified Dulbecco’s medium (IMDM) (Euroclone) containing 10% heat-inactivated fetal bovine serum (Euroclone), 100 IU of penicillin, streptomycin (100μgmL−1), 2 mM l-glutamine (Euroclone), and granulocyte-macrophage colony-stimulating factor (GM-CSF) (10 to 20ngmL–1) for 8 days.
Colombo M., Marongiu L., Mingozzi F., Marzi R., Cigni C., Facchini F.A., Rotem R., Valache M., Stucchi G., Rocca G., Gornati L., Rizzuto M.A., Salvioni L., Zanoni I., Gori A., Prosperi D, & Granucci F. (2022). Specific immunosuppressive role of nanodrugs targeting calcineurin in innate myeloid cells. iScience, 25(10), 105042.
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6

Curcumin's Effects on Cell Apoptosis and Proliferation

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HEL cells were incubated with different concentrations (10, 15, 20, 30 µmol/L) of curcumin (stock solution 50 mmol/L in DMSO, #C1386, Sigma‐Aldrich) for 24 and 48 hours. Leucocytes isolated from MPNs patients were incubated with 30 µmol/L of curcumin in IMDM (EuroClone) supplemented with 20% inactivated FBS for 20 hours. After incubation, apoptosis and proliferation were evaluated and total RNA and proteins were extracted as described below.
Petiti J., Rosso V., Lo Iacono M., Panuzzo C., Calabrese C., Signorino E., Pironi L., Cartellà A., Bracco E., Pergolizzi B., Beltramo T., Fava C, & Cilloni D. (2019). Curcumin induces apoptosis in JAK2‐mutated cells by the inhibition of JAK2/STAT and mTORC1 pathways. Journal of Cellular and Molecular Medicine, 23(6), 4349-4357.
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7

Isolation and Cultivation of CD34+ Cells for PMF

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Human CD34+ cells were purified from CB or PB samples from PMF patients as previously described [21 (link)]. Purified CD34+ cells were cultured in 24-well plates at 5×105/ml in Iscove's modified Dulbecco's medium (IMDM, Euroclone) containing 20% HS (Bio-Whittaker), interleukin-3 (IL-3) (10 ng/ml), interleukin-6 (IL-6) (10 ng/ml), thrombopoietin (TPO) (20 ng/ml), stem cell factor (SCF) (50 ng/ml) and Flt3-ligand (FLT3L) (50 ng/ml), (all from Miltenyi Biotec; Auburn, CA, USA) and electroporated 24 hours later.
Rontauroli S., Norfo R., Pennucci V., Zini R., Ruberti S., Bianchi E., Salati S., Prudente Z., Rossi C., Rosti V., Guglielmelli P., Barosi G., Vannucchi A., Tagliafico E, & Manfredini R. (2017). miR-494-3p overexpression promotes megakaryocytopoiesis in primary myelofibrosis hematopoietic stem/progenitor cells by targeting SOCS6. Oncotarget, 8(13), 21380-21397.
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8

Cell Culture and Authentication of EWS Lines

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EWS cell lines were grown as previously described13 (link),40 (link). The lines 6647 and TC-71 were kindly provided by T.J. Triche (Children's Hospital, Los Angeles, CA, USA); SK-N-MC, SK-ES-1, and RD-ES were provided by American Type Culture Collection, ATCC (Rockville, MD, USA); IOR/CAR and LAP-35 were previously established in our laboratory; and the A673 sarcoma cell line was provided by Dr. H. Kovar (St. Anna Kinderkrebsforschung, Vienna Austria). Stable CD99-silenced cells were obtained from the TC-71 and IOR/CAR cell lines as previously described12 (link),13 (link). The cells were cultured in Iscove's modified Dulbecco's medium (IMDM; EuroClone, Milan, Italy) enriched with 10% fetal bovine serum (FBS) (EuroClone) supplemented with 100 U/ml penicillin and 100 mg/ml streptomycin and incubated at 37 °C in a humidified atmosphere at 5% CO2.
All cell lines were tested for mycoplasma contamination (MycoAlert Mycoplasma Detection Kit, Lonza) and authenticated by short tandem repeat (STR) polymerase chain reaction (PCR) analysis (last control: December 2017) using a PowerPlex ESX Fast System kit (Promega, Madison, WI, USA).
De Feo A., Sciandra M., Ferracin M., Felicetti F., Astolfi A., Pignochino Y., Picci P., Carè A, & Scotlandi K. (2019). Exosomes from CD99-deprived Ewing sarcoma cells reverse tumor malignancy by inhibiting cell migration and promoting neural differentiation. Cell Death & Disease, 10(7), 471.
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9

Culturing Human Dermal Fibroblasts

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Human dermal fibroblasts CCD-1064Sk (ATCC® CRL-2076TM, Manassas, VA, USA) were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM, Euroclone, Pero MI, Italy) supplemented with 10% v/v fetal bovine serum (FBS, Euroclone, Pero MI, Italy) and 1% v/v of 200 mM L-glutamine. Penicillin-Streptomycin; 100 IU/mL–100 µg/mL (Euroclone, Pero MI, Italy) was added to the media. Cells were kept at 37 °C in a humidified atmosphere containing 5% CO2 and used at a confluence of 80–90%.
Mahmoud N.N., Al-Kharabsheh L.M., Khalil E.A, & Abu-Dahab R. (2019). Interaction of Gold Nanorods with Human Dermal Fibroblasts: Cytotoxicity, Cellular Uptake, and Wound Healing. Nanomaterials, 9(8), 1131.
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10

Culturing of Gastric Cancer Cell Lines

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The human HER2+ GC cell lines OE19, N87, OE33 and the human HER2-negative GC cell line MKN45 were obtained from ATCC (Rockville, MD, USA) and grown as monolayer cultures in RPMI 1640 (EuroClone, Pero, MI, Italy) with 10% FBS. GC primary cells GTR0455 were derived from PDX as described in [24 ] and grown as a monolayer culture in IMDM (EuroClone) with 10% FBS. All tumor cell lines were cultured at 37 °C in a humidified 5% CO2 atmosphere and routinely tested for mycoplasma contamination.
Castagnoli L., Corso S., Franceschini A., Raimondi A., Bellomo S.E., Dugo M., Morano F., Prisciandaro M., Brich S., Belfiore A., Vingiani A., Di Bartolomeo M., Pruneri G., Tagliabue E., Giordano S., Pietrantonio F, & Pupa S.M. (2023). Fatty acid synthase as a new therapeutic target for HER2-positive gastric cancer. Cellular Oncology (Dordrecht), 46(3), 661-676.
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