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Ficoll separation

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Ficoll separation is a laboratory technique used to isolate specific cell types from a heterogeneous mixture. It utilizes a density gradient centrifugation process to separate different components based on their densities.

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Lab products found in correlation

Gm csf, by Thermo Fisher Scientific (2 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Rosettesep monocyte enrichment kit, by STEMCELL (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Interleukin 4 (il 4), by R&D Systems (1 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) X vivo 15 medium, by Lonza (1 mentions)

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Ficoll

3 protocols using ficoll separation

1

Generation of Human Monocyte-Derived Dendritic Cells

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Human Buffy coats were acquired from Karolinska Institutet, Stockholm, Sweden. All experimental work with human peripheral blood cells were carried out in accordance with Swedish guidelines and regulations. All experimental protocols were approved by the local ethical committee in Stockholm (“Regionala etikprövningsnämnden i Stockholm”, Ethical permit Dnr 2006/229-31/3). According to regulations in Sweden, experimental in vitro work with cells from buffy coats does not require informed consent. Human monocytes were generated using the RosetteSep Monocyte Enrichment Kit (1 mL/10 mL buffy coat; StemCell Technologies) and differentiated into moDC, with GM-CSF (250 ng/mL; PeproTech) and IL-4 (6.5 ng/mL; R&D Systems) for 6 days in +37 °C, 5% CO2 at a density of 5 × 105 cells/mL in RPMI 1640 completed with 10% FCS, 1 mM sodium pyruvate, 10 mM HEPES, 2mM L-glutamine, and 1% streptomycin and penicillin (all from Invitrogen Life Technologies) as previously described17 (link). Differentiation of the cells was monitored by CD1a expression. PBMCs were isolated from buffy coats after Ficoll separation (Stemcell).
Järver P., Dondalska A., Poux C., Sandberg A., Bergenstråhle J., Sköld A.E., Dereuddre-Bosquet N., Martinon F., Pålsson S., Zaghloul E., Brodin D., Sander B., Lennox K.A., Behlke M.A., EL-Andaloussi S., Lehtiö J., Lundeberg J., LeGrand R, & Spetz A.L. (2018). Single-Stranded Nucleic Acids Regulate TLR3/4/7 Activation through Interference with Clathrin-Mediated Endocytosis. Scientific Reports, 8, 15841.
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2

Isolation of Peripheral Blood Mononuclear Cells

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30 to 50 ml of peripheral blood was drawn into heparinized tubes and peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll separation (Stem Cell Technologies) as previously described (6 (link)).
Sun H., Zhu Y., Pan H., Chen X., Balestrini J.L., Lam T.T., Kanyo J.E., Eichmann A., Gulati M., Fares W.H., Bai H., Feghali-Bostwick C.A., Gan Y., Peng X., Moore M.W., White E.S., Sava P., Gonzalez A.L., Cheng Y., Niklason L.E, & Herzog E.L. (2016). Netrin-1 regulates fibrocyte accumulation in the decellularized fibrotic scleroderma lung microenvironment and in bleomycin induced pulmonary fibrosis. Arthritis & rheumatology (Hoboken, N.J.), 68(5), 1251-1261.
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3

Monocyte Isolation and Differentiation

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Monocytes were enriched from peripheral blood mononuclear cells from three healthy donors using Rossette Sep Monocyte enrichment cocktail through Ficoll separation (STEMCELL Technologies, Vancouver, Canada) as previously described (22 (link)). Purified monocytes were then differentiated in X-vivo15 medium (Lonza, Walkersville, MD, USA), with GMCSF 1000U/ml and IL4 1000U/ml (Pepro Tech Inc, Rocky Hill, NJ, USA) with the change of media every other day.
Elsayed R., Kurago Z., Cutler C.W., Arce R., Gerber J., Celis E., Sultan H., Elashiry M., Meghil M., Sun C., Auersvald C.M., Elsayed R., Elshikh M.E., Isales C, & Elsalanty M.E. (2020). Role of dendritic cell - mediated immune response in oral homeostasis: a new mechanism of osteonecrosis of the jaw. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(2), 2595-2608.
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