Roswell park memorial institute (rpmi)

Manufactured by R&D Systems

Sourced in United States

RPMI is a commonly used cell culture medium formulation. It provides a balanced salt solution, amino acids, vitamins, and other nutrients to support the growth and maintenance of various cell types in vitro.

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Lab products found in correlation

Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (3 mentions) Recombinant rat igf 1, by R&D Systems (1 mentions) Charcoal stripped fbs, by Thermo Fisher Scientific (1 mentions) Cell proliferation kit 1, by Roche (1 mentions) Human ab serum, by Merck Group (1 mentions) Roswell park memorial institute (rpmi), by Merck Group (1 mentions) Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions) M csf, by R&D Systems (1 mentions) Pancoll human, by PAN Biotech (1 mentions) Bone morphogenetic protein 4 (bmp4), by Thermo Fisher Scientific (1 mentions) Fibroblast growth factor 2 (fgf2), by Thermo Fisher Scientific (1 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions) Oncostatin m, by R&D Systems (1 mentions) Activin a, by R&D Systems (1 mentions) Bone morphogenetic protein 4 (bmp4), by R&D Systems (1 mentions) Growth factor reduced matrigel, by BD (1 mentions) Hepatocyte growth factor (hgf), by R&D Systems (1 mentions) Transwell insert, by Corning (1 mentions) Hepes, by Merck Group (1 mentions) Fura 2, by Thermo Fisher Scientific (1 mentions)

5 protocols using roswell park memorial institute (rpmi)

Evaluating IGF-1 and DHT Effects on Dunning G Cells

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Dunning G cells (5000 cells/well) were seeded in 100 µl phenol-free RPMI (Gibco) supplemented with 2.5% charcoal stripped FBS (Gibco), in 96-well plates, and allowed to settle overnight. Culture medium was carefully removed and replaced with 100 µl phenol-free RPMI supplemented with 2.5% charcoal stripped FBS and various concentrations of recombinant rat IGF-1 (R&D Systems, Minneapolis, MN, US), dihydrotestosterone (DHT, Sigma-Aldrich, St Louis, MO, US) and NVP-AEW541, at doses previously described [13 (link)]. The relative amount of viable tumor cells was measured at baseline, and day 2, 4 and 7 using the Cell Proliferation Kit I (Roche Diagnostics, Bromma, Sweden) according to the manufacturer’s instructions. Cells that were cultured for 7 days received fresh media with supplements at day 4 of the experiment.

Nordstrand A., Bergström S.H., Thysell E., Bovinder-Ylitalo E., Lerner U.H., Widmark A., Bergh A, & Wikström P. (2017). Inhibition of the insulin-like growth factor-1 receptor potentiates acute effects of castration in a rat model for prostate cancer growth in bone. Clinical & Experimental Metastasis, 34(3), 261-271.

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Isolation and Differentiation of Human Monocyte-Derived Macrophages

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Human blood samples were obtained from healthy donors (16 donors, 5 females and 11 males, aged 23 to 50 years with a median age of 30 years). Heparinized blood was centrifuged using a density-gradient (Pancoll Human, PAN Biotech, Aidenbach, Germany). The contained peripheral blood mononuclear cells (PBMC) were retrieved and washed using Hanks buffered saline solution (HBSS) (Gibco, UK). PBMCs were re-suspended in RPMI (Sigma, Saint Louis, USA) with 5% AB human serum (Sigma) and evenly seeded into six-well plates. After two hours of incubation at 37°C with 5% CO_2, the remaining non-adherent cells were removed by gently washing the plates with HBSS three times. The adherent monocytes were then incubated for three days in RPMI with 10% autologous human serum and 20 ng/ml macrophage-colony stimulating factor (M-CSF) (R&D Systems, Minneapolis, USA). The resident cells were then again washed with HBSS and incubated for three days in RPMI with 10% autologous serum until the differentiation to MDM was completed [15 (link)].

Kammrath Betancor P., Hildebrand A., Böhringer D., Emmerich F., Schlunck G., Reinhard T, & Lapp T. (2018). Activation of human macrophages by human corneal allogen in vitro. PLoS ONE, 13(4), e0194855.

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Directed Differentiation of iPSCs into iHeps

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Directed differentiation of iPScs into iHeps was performed in vitro using a variation of the 4-step protocol described by Si-Tayeb.^{16 (link)} Briefly, iPScs were single-cell passaged onto growth factor reduced Matrigel™ (BD Biosciences, San Jose, CA) and induced to differentiate into definitive endoderm cells by treatment with RPMI (Invitrogen, Carlsbad, CA), 1X B27 w/o insulin (Invitrogen, Carlsbad, CA) and 0.5X NEAA (Invitrogen, Carlsbad, CA) containing 100 ng/mL activin A (R&D Systems, Minneapolis, MN), 10 ng/mL BMP4 (R&D Systems, Minneapolis, MN) and 20 ng/mL FGF2 (Biosciences, San Jose, CA) for 2 days followed by treatment with RPMI, 1X B27 w/o insulin and 0.5X NEAA with 100 ng/mL activin A for 3 days at ambient O₂/5% CO₂. For hepatic specification, the cells were cultured in RPMI, 1X B27 w/insulin (Invitrogen, Carlsbad, CA) and 0.5X NEAA containing 20 ng/mL BMP4 and 10 ng/mL FGF2 for 5 days at 4% O₂/5% CO₂. For hepatic induction, the cells were treated with RPMI, 1X B27 w/insulin and 0.5X NEAA containing 20 ng/mL HGF (R&D Systems, Minneapolis, MN) for 5 days at 4% O₂/5% CO₂. For hepatic maturation, the cells were cultured for 5 days in HCM™ (Lonza, Walkersville, MD) without EGF and with 20 ng/mL oncostatin M (R&D Systems, Minneapolis, MN) at ambient O₂/5% CO₂.

Tafaleng E.N., Chakraborty S., Han B., Hale P., Wu W., Soto-Gutierrez A., Feghali-Bostwick C.A., Wilson A.A., Kotton D.N., Nagaya M., Strom S.C., Chowdhury J.R., Stolz D.B., Perlmutter D.H, & Fox I.J. (2015). Induced pluripotent stem cells model personalized variations in liver disease due to α1-antitrypsin deficiency. Hepatology (Baltimore, Md.), 62(1), 147-157.

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Transwell Monocyte Migration Assay

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Isolated blood cells were washed in RPMI (Gibco) medium containing 10 mM HEPES (Sigma) and 0.1% bovine serum albumin (pH 7.4) before being transferred into a Transwell insert (3 × 10⁵cells/well, Costar Corning) previously coated with recombinant human Fc‐tagged murine VCAM (5 μg/ml, R&D systems) in PBS. The inserts were then placed on a 24‐well plate filled with RPMI medium (10 mM HEPES, 0.1% BSA, pH 7.4), with or without stimulation of mCCL2 (20 ng/ml, R&D systems). After 2 hr incubation at 37 °C and 5% CO₂, cells were isolated from the lower compartment and counted under a light microscope.

Düsedau H.P., Kleveman J., Figueiredo C.A., Biswas A., Steffen J., Kliche S., Haak S., Zagrebelsky M., Korte M, & Dunay I.R. (2018). p75NTR regulates brain mononuclear cell function and neuronal structure in Toxoplasma infection‐induced neuroinflammation. Glia, 67(1), 193-211.

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Fura2-Based Ca2+ Imaging of EPO-Treated HPCs

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A fluorescence microscopy-coupled digital imaging system was used to measure Ca2+ concentration change of HPCs when treated with EPO. Fura2 was used as the detection fluorophore. After the whole yolk sacs were dissociated into single cells, the cells were washed 2-3 times with PBS to completely remove fetal bovine serum (FBS). HPCs were resuspended in 40ul RPMI 1640 (Sigma) without FBS. Cells were loaded into the center of culture dish coated with poly-lysine and fixed for 10min at room temperature. The medium was then removed, and cells were stained with 1µM Fura2 and Pluronic F127 (Invitrogen) (1:1 in volume) at 37°C for 30min.
Staining solution was removed, and cells were covered with 150µl RPMI containing 10% FBS.
Mouse EPO (R&D systems) was added to a final concentration of 10 units/ml. Then the sample was analyzed immediately with the confocal fluorescence microscopy on a Nikon TE2000U microscope. Fura2-loaded cells were visualized with digital video imaging, and fluorescence was quantitated using the intensity ratio of the emission (510nm), which was measured following excitation at 340nm divided by the emission following excitation at 380nm. Each sample was measured over 20min. After that, the data was calculated and analyzed by Metamorph 6.1 (Universal Imaging Corp., Downingtown, PA).

Feng Y., Borosha S., Ratri A., Housami S.M., Chakravarthi V.P., Wang H., Vivian J.L., Fields T.A., Kinsey W.H., Rumi M.A.K., & Fields P.E. (2020). DOT1L methyltransferase regulates the calcium influx in erythroid progenitor cells in response to erythropoietin.

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