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Ficoll histopaque solution

Manufactured by Merck Group
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Sourced in Brazil, United States

Ficoll-Histopaque solution is a density gradient medium used for the separation and purification of cells, such as lymphocytes, from whole blood or other biological samples. It is a sterile, endotoxin-tested solution composed of a mixture of Ficoll and sodium diatrizoate.

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Lab products found in correlation

Lsr 2 flow cytometer, by BD (1 mentions) Facsdiva software, by BD (1 mentions) Ficoll histopaque, by Merck Group (1 mentions) Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Tripure reagent, by Roche (1 mentions) Cd14 mab coated micro beads, by Miltenyi Biotec (1 mentions)

Close spelling variants of this product

Histopaque (690 citations) Ficoll (251 citations) Ficoll-Histopaque (184 citations) Histopaque solution (31 citations) Ficoll solution (16 citations)

4 protocols using ficoll histopaque solution

1

Phenotypic Analysis of PBMCs

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The blood and lymph node aspirates were collected before and after infection. Briefly, PBMCs were isolated from 10ml of blood using Ficoll Histopaque solution (Sigma-Aldrich). Then, cells were stained with CD3, CD4, CD8, CD19, CD40, CD11c and CD86 antibodies and isotype control. Data collection and analysis were performed using a BD LSR-II flow cytometer, BD FACSDiva software (BD Pharmingen), and FlowJo software (TreeStar, San Carlos, CA).
Gnanadurai C.W., Yang Y., Huang Y., Li Z., Leyson C.M., Cooper T.L., Platt S.R., Harvey S.B., Hooper D.C., Faber M, & Fu Z.F. (2015). Differential Host Immune Responses after Infection with Wild-Type or Lab-Attenuated Rabies Viruses in Dogs. PLoS Neglected Tropical Diseases, 9(8), e0004023.
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2

Expansion and Engineering of CB-NK Cells

Cited 1 time
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CB units for research were provided by the MD Anderson Cancer Center (MDACC) CB Bank under an IRB approved protocol (Lab04-0249). CB-derived NK cells (CB-NK) were isolated and expanded as previously described36 (link). In brief, lymphocytes were collected by density-gradient centrifugation using Ficoll-Histopaque solution (Sigma-Aldrich). CD56+CD3− NK cells were then purified using an NK negative isolation kit (Miltenyi Biotec), and co-cultured with irradiated (100 Gy) uAPCs at a 2:1 ratio in complete stem cell growth medium (SCGM), supplemented with 200 U/ml recombinant human IL-2 (Proleukin). On Day 4 post uAPC stimulation, fresh NK cells were purified again and transduced with retroviral vectors expressing CAR-constructs. A second retroviral transduction of iCAR constructs was performed on Day 6 to then generate NK cells expressing AI-CAR. Following the same approach, CD19-mCherry or CD19-mCherry/GFP expressing cells (both primary NK cells and tumor cells) were prepared. CAR transduction efficiency was measured by flow cytometry. Irradiated uAPC were added weekly to the NK cell culture to support NK cell expansion.
Li Y., Basar R., Wang G., Liu E., Moyes J.S., Li L., Kerbauy L.N., Uprety N., Fathi M., Rezvan A., Banerjee P.P., Muniz-Feliciano L., Laskowski T.J., Ensley E., Daher M., Shanley M., Mendt M., Acharya S., Liu B., Biederstädt A., Rafei H., Guo X., Garcia L.M., Lin P., Ang S., Marin D., Chen K., Bover L., Champlin R.E., Varadarajan N., Shpall E.J, & Rezvani K. (2022). KIR-based inhibitory CARs overcome CAR-NK cell trogocytosis-mediated fratricide and tumor escape. Nature medicine, 28(10), 2133-2144.
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3

Isolation and Cultivation of Bovine PBMCs

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Blood samples were collected from adult Holstein cows in a rural area of Bahia, Brazil. Bovine peripheral blood was collected from the jugular vein into vacutainer tubes containing 1.5 mg/mL EDTA (BD Vacutainer®) and diluted at 1:1 in PBS (pH 7.4). Next, 10 mL of the mixture was added to 3 mL of Ficoll–Histopaque solution (density: 1.0771 g/mL, Sigma-Aldrich, São Paulo, Brazil) in a 15 mL tube to produce the Ficoll–Histopaque barrier, followed by centrifugation at 4200× g for 20 min at room temperature. The mononuclear cells present at the Ficoll/plasma interface were removed, washed twice in PBS, and centrifuged at 4200× g for 10 min after each wash step. PBMCs were counted in a Neubauer chamber, and cell viability was assessed using 0.1% trypan blue (viability > 90%). Viable cell concentrations were adjusted to 1 × 106 cells/mL by adding Roswell Park Memorial Institute (RPMI)-1640 medium supplemented with 10% foetal bovine serum (FBS; Gibco, São Paulo, Brazil) and cultured in complete RPMI-1640 medium supplemented with 10% FBS and 100 U/mL penicillin.
Santos-Junior M.N., Neves W.S., Santos R.S., Almeida P.P., Fernandes J.M., Guimarães B.C., Barbosa M.S., da Silva L.S., Gomes C.P., Sampaio B.A., Rezende I.D., Correia T.M., Neres N.S., Campos G.B., Bastos B.L., Timenetsky J, & Marques L.M. (2022). Heterologous Expression, Purification, and Immunomodulatory Effects of Recombinant Lipoprotein GUDIV-103 Isolated from Ureaplasma diversum. Microorganisms, 10(5), 1032.
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4

Isolation and Purification of Monocytes

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Whole blood collected in EDTA tubes was diluted 1:1 with 1X PBS–1% heparin and subsequently added to a Ficoll-Histopaque solution (Sigma, Sigma, St. Louis, MO, USA, Catalog Number 10771). The samples were centrifuged at 1300 rpm for 30 min, and the peripheral blood mononuclear cells (PBMCs) were generated after the centrifugation. Then, positive selection to obtain monocytes was carried out using CD14-mAb-coated microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany; Catalog Number 130-050-201) following the manufacturer’s instructions (purity of 95 ± 98%). An aliquot of 1 mL of Tripure TM reagent (Roche Life Science, Penzberg, Germany) was added for collecting the monocytes. The cells were stored at −80 °C.
Torres-Paz Y.E., Gamboa R., Fuentevilla-Álvarez G., Soto M.E., González-Moyotl N., Martínez-Alvarado R., Torres-Tamayo M., Ramírez-Marroquín E.S., Vásquez-Jiménez X., Sainz-Escarrega V, & Huesca-Gómez C. (2023). Overexpression of microRNA-21-5p and microRNA-221-5p in Monocytes Increases the Risk of Developing Coronary Artery Disease. International Journal of Molecular Sciences, 24(10), 8641.
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