Iscove s mdm

Manufactured by Thermo Fisher Scientific

Iscove's MDM is a complete, chemically defined medium designed for the culture of a variety of cell types, including hematopoietic, myeloid, and lymphoid cells. It provides the necessary nutrients and growth factors to support the in vitro growth and maintenance of these cell types.

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Lab products found in correlation

Ant pr, by InvivoGen (1 mentions) Es grade fbs, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) Rmpi 1640, by Thermo Fisher Scientific (1 mentions) Bovine pancreatic insulin, by Merck Group (1 mentions) Human transferrin, by Merck Group (1 mentions) Hi fbs, by Merck Group (1 mentions) Ril 3, by R&D Systems (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) 2 mercaptoethanol, by Merck Group (1 mentions) Methylcellulose, by Merck Group (1 mentions)

4 protocols using iscove s mdm

Culture and Maintenance of Murine G1ER Cells

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The murine G1ER cell line was kindly provided by Drs. Mitchell Weiss and Stuart Orkin.^{38 (link)–40 (link)} Cells were maintained between 1 × 10⁵ and 2 × 10⁶ cells/mL by dilution into fresh medium or by centrifuging the cells at 125 g × 5 min and resuspending in fresh medium. G1ER cells were cultured in media containing 500 mL Iscove’s MDM (Gibco, 12440053), 75 mL ES-grade FBS (Gibco, 16141079), 10 mL penicillin/streptomycin stock (Gibco 15140122), 6.2 μL monothioglycerol (MTG) (Sigma, M6145), 100 μL erythropoietin (Procrit, 10,000 U/mL stock), and 50 μL puromycin (InvivoGen, ant-pr; ~1 μg/mL). Fresh mouse stem cell factor was added when passaging cells at 20 ng / mL with a 1000X dilution (Sigma S9915, resuspended in 0.8X PBS).

Dolberg T.B., Gunnels T.F., Ling T., Sarnese K.A., Crispino J.D, & Leonard J.N. (2023). Building synthetic biosensors using red blood cell proteins. bioRxiv.

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Culturing Hematopoietic Cell Lines

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K562, K562-A1, FM6, FM28, and FM82 cell lines were maintained in RMPI-1640 (Gibco) supplemented with 10% FBS. OCI-M2 was maintained in Iscove’s MDM (Gibco) with 20% FBS. Cells were passaged every 2–3 days. Adherent cells (FM6, FM28, and FM82) were passaged following detachment from the flask with 0.25% trypsin (Gibco). All cells lines were confirmed to be mycoplasma negative.

Weis-Banke S.E., Lisle T.L., Perez-Penco M., Schina A., Hübbe M.L., Siersbæk M., Holmström M.O., Jørgensen M.A., Marie Svane I., Met Ö., Ødum N., Madsen D.H., Donia M., Grøntved L, & Andersen M.H. (2022). Arginase-2-specific cytotoxic T cells specifically recognize functional regulatory T cells. Journal for Immunotherapy of Cancer, 10(10), e005326.

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GEMM-CFU Assay for Bone Marrow Cells

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GEMM-CFU were assayed using a complete growth media/methylcellulose (Sigma-Aldrich) based colony assay (Cortdy 1995 , Seed et al. 2002a ). The complete medium was comprised of: methylcellulose (Sigma-Aldrich) in Iscove’s MDM (Life Technologies), HI-FBS, bovine serum albumin (Sigma-Aldrich), bovine pancreatic insulin (Sigma-Aldrich), human transferrin (Sigma-Aldrich), 2-mercaptoethanol (Sigma- Aldrich), L-glutamine (Life Technologies), recombinant stem cell factor (rSCF; Pharmingen), recombinant interleukin-3 (rIL-3; R&D Systems, Minneapolis, MN), recombinant human interleukin-6 (rhIL-6; R&D Systems), and recombinant human erythropoietin (rh erythropoietin, R&D Systems). Nucleated bone marrow cells were diluted to 1.5 × 10⁵ cells/ml in IMDM with 2% HI-FBS; 0.3 ml of diluted cells were mixed with 3.0 ml of the complete medium. Cell/media mix (1 ml) was dispensed to each 35 mm plate. GEMM-CFU colonies showing all four lineages were scored 14 days after incubation in a 37°C humidified environment containing 5% CO₂.

Seed T.M., Inal C.E, & Singh V.K. (2014). Radioprotection of hematopoietic progenitors by low dose amifostine prophylaxis. International Journal of Radiation Biology, 90(7), 594-604.

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Humanized Xenograft Models for Leukemia

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NSG xenograft recipients (6–8-week old) were used with IACUC approval. Conditioned Molm-14 cells (1 × 10⁵), cord-blood CD34+ cells or 5 × 10⁶ HL-60 cells were resuspended in Hank's balanced salt solution media and injected via tail vein. Hank's balanced salt solution medium was used as vehicle control in all xenograft experiments. Human CD45 chimerism (BioLegend, HI30, San Diego, CA, USA) was monitored by flow cytometry. Animals were killed at 3–5-weeks post engraftment, and peripheral blood (PB) and BM were collected. Adherent BM stromal cells were propagated in Iscove's MDM (Life Technologies) with 10% VF FBS (detailed description in Supplementary Materials and Methods).

Huan J., Hornick N., Goloviznina N., Kamimae- Lanning A., David L., Wilmarth P., Mori T., Chevillet J., Narla A., Roberts C J.r., Loriaux M., Chang B, & Kurre P. (2015). Coordinate regulation of residual bone marrow function by paracrine trafficking of AML exosomes. Leukemia, 29(12), 2285-2295.

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