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Ficoll paque plus 206

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Ficoll Paque™ Plus 206 is a density gradient medium used for the separation and purification of cells, organelles, and other biological particles. It is designed to provide efficient and gentle separation of different cell types, such as lymphocytes, monocytes, and granulocytes, from whole blood or other complex biological samples.

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Lab products found in correlation

Facsymphony a5, by BD (1 mentions) Rpmi 1640 medium, by Euroclone (1 mentions) Facslyric, by BD (1 mentions) Pharm lyse, by BD (1 mentions)

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Ficoll-Paque Plus (419 citations) Ficoll-Paque (225 citations) Ficoll (53 citations)

8 protocols using ficoll paque plus 206

1

Phenotypic Analysis of PBMC Subsets

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Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll Paque™ Plus 206 (Amersham Pharmacia Biotech, Amersham, UK) density-gradient centrifugation and immediately frozen and stored in liquid nitrogen until use. The freezing medium contained 90% Fetal Bovine Serum (FBS) and 10% DMSO. Cells were stained with the appropriate combination of fluorochrome-conjugated antibodies to identify T-cell and B-cell subsets according to standard techniques. B-cell populations were identified as CD19+. Memory B cells (MBC) were gated as CD19+CD24+CD27+, and IgM and switched memory B cells were discriminated by the expression of IgM. T cell subsets were identified based on the expression of CD3, CD4, CD8, CD45RO and CCR7 markers by flow cytometry. An analysis of T cell subsets was performed in all 47 CVID patients and in 9 age-matched HD.
Immune cells were identified as follows: naïve CD4+ T cells (CD4+CD45RO−CCR7+), central memory CD4+ (CD4+CD45RO+CCR7−) T cells, naïve CD8+ T cells (CD8+CD45RO-) and central memory CD8+ T cells (CD8+CD45RO+CCR7−). The CD4+ naïve/memory ratio was calculated as a ratio of CD4+CD45RO−CCR7+ and CD4+CD45RO+CCR7−; the CD8+ naïve/memory ratio was calculated as a ratio of CD8+CD45RO−CCR7+ and CD8+CD45RO+CCR7−. Cells were acquired on a BD FACSymphony A5™. Data were analyzed with FlowJo ver. 10.
Pulvirenti F., Di Cecca S., Sinibaldi M., Piano Mortari E., Terreri S., Albano C., Guercio M., Sculco E., Milito C., Ferrari S., Locatelli F., Quintarelli C., Carsetti R, & Quinti I. (2022). T-Cell Defects Associated to Lack of Spike-Specific Antibodies after BNT162b2 Full Immunization Followed by a Booster Dose in Patients with Common Variable Immune Deficiencies. Cells, 11(12), 1918.
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2

PBMC Isolation and Cryopreservation

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Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll Paque™ Plus 206 (Amersham PharmaciaBiotech) density-gradient centrifugation and immediately frozen and stored in liquid nitrogen until use. The freezing medium contained 90% Fetal Bovine Serum (FBS) and 10% DMSO.
Piano Mortari E., Pulvirenti F., Marcellini V., Terreri S., Salinas A.F., Ferrari S., Di Napoli G., Guadagnolo D., Sculco E., Albano C., Guercio M., Di Cecca S., Milito C., Garzi G., Pesce A.M., Bonanni L., Sinibaldi M., Bordoni V., Di Cecilia S., Accordini S., Castilletti C., Agrati C., Quintarelli C., Zaffina S., Locatelli F., Carsetti R, & Quinti I. (2023). Functional CVIDs phenotype clusters identified by the integration of immune parameters after BNT162b2 boosters. Frontiers in Immunology, 14, 1194225.
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3

PBMC Isolation and Cryopreservation

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Heparinized PBMCs were isolated by Ficoll Paque™ Plus 206 (Amersham PharmaciaBiotech, Amersham, UK) density-gradient centrifugation and immediately frozen and stored in liquid nitrogen until use. The freezing medium contained 90% Foetal Bovine Serum (FBS) and 10% DMSO.
Piano Mortari E., Russo C., Vinci M.R., Terreri S., Fernandez Salinas A., Piccioni L., Alteri C., Colagrossi L., Coltella L., Ranno S., Linardos G., Agosta M., Albano C., Agrati C., Castilletti C., Meschi S., Romania P., Roscilli G., Pavoni E., Camisa V., Santoro A., Brugaletta R., Magnavita N., Ruggiero A., Cotugno N., Amodio D., Ciofi Degli Atti M.L., Giorgio D., Russo N., Salvatori G., Corsetti T., Locatelli F., Perno C.F., Zaffina S, & Carsetti R. (2021). Highly Specific Memory B Cells Generation after the 2nd Dose of BNT162b2 Vaccine Compensate for the Decline of Serum Antibodies and Absence of Mucosal IgA. Cells, 10(10), 2541.
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4

PBMC Isolation and Cryopreservation

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Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll Paque Plus 206 (Amersham PharmaciaBiotech) density-gradient centrifugation and immediately frozen and stored in liquid nitrogen until use. The freezing medium contained 90% Fetal Bovine Serum (FBS) and 10% dimethyl sulfoxide (DMSO).
Pulvirenti F., Mortari E.P., Putotto C., Terreri S., Fernandez Salinas A., Cinicola B.L., Cimini E., Di Napoli G., Sculco E., Milito C., Versacci P., Agrati C., Marino B., Carsetti R, & Quinti I. (2022). COVID-19 Severity, Cardiological Outcome, and Immunogenicity of mRNA Vaccine on Adult Patients With 22q11.2 DS. The Journal of Allergy and Clinical Immunology. in Practice, 11(1), 292-305.e2.
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5

Isolation and Cryopreservation of PBMCs

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Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll Paque™ Plus 206 (Amersham PharmaciaBiotech, Amersham, UK) density-gradient centrifugation and immediately frozen and stored in liquid nitrogen until use.
Cinicola B.L., Piano Mortari E., Zicari A.M., Agrati C., Bordoni V., Albano C., Fedele G., Schiavoni I., Leone P., Fiore S., Capponi M., Conti M.G., Petrarca L., Stefanelli P., Spalice A., Midulla F., Palamara A.T., Quinti I., Locatelli F, & Carsetti R. (2022). The BNT162b2 vaccine induces humoral and cellular immune memory to SARS-CoV-2 Wuhan strain and the Omicron variant in children 5 to 11 years of age. Frontiers in Immunology, 13, 1094727.
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6

PBMC Isolation and Cryopreservation

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Heparinized peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll Paque™ Plus 206 (Amersham PharmaciaBiotech) density-gradient centrifugation and immediately frozen and stored in liquid nitrogen until use. The freezing medium contained 90% Fetal Bovine Serum (FBS) and 10% DMSO.
Salinas A.F., Mortari E.P., Terreri S., Quintarelli C., Pulvirenti F., Di Cecca S., Guercio M., Milito C., Bonanni L., Auria S., Romaggioli L., Cusano G., Albano C., Zaffina S., Perno C.F., Spadaro G., Locatelli F., Carsetti R, & Quinti I. (2021). SARS-CoV-2 Vaccine Induced Atypical Immune Responses in Antibody Defects: Everybody Does their Best. Journal of Clinical Immunology, 41(8), 1709-1722.
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7

PBMC Isolation from Heparinized Blood

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Heparinized peripheral blood was collected and then centrifuged through a Histopaque gradient (Ficoll Paque™ Plus 206, Amersham Pharmacia Biotech) for 25 min at 2400 rpm. PBMCs were collected from the interface. Next, PBMCs were spun down and washed twice with RPMI 1640 medium (Euroclone). Then PBMCs were frozen and stored in liquid nitrogen until use. The freezing medium contained 90% Fetal Bovine Serum (FBS) and 10% DMSO.
Ciocca M., Zaffina S., Fernandez Salinas A., Bocci C., Palomba P., Conti M.G., Terreri S., Frisullo G., Giorda E., Scarsella M., Brugaletta R., Vinci M.R., Magnavita N., Carsetti R, & Piano Mortari E. (2021). Evolution of Human Memory B Cells From Childhood to Old Age. Frontiers in Immunology, 12, 690534.
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8

Multiparameter Leukocyte Profiling

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Four leukocyte profiling panels computing seven- to nine-surface marker antigens for monitoring the major leukocyte subsets as well as characteristics of T cell, B cell, monocytes and NK cells subsets were designed (Supplementary Table S2). Results of immune-profile of analyzed patients are reported in Supplementary Tables S3–10. The graphs of the single time points refer to the sample obtained immediately after the first positive nasopharyngeal swab. When available, we also show the results obtained at different time points.
1 ml of total blood (EDTA) was incubated with the lysing solution Pharm Lyse (BD) to lyse red blood cells. Then, cells were divided in four equal aliquots and stained with the appropriate combination of fluorochrome-conjugated antibodies (Supplementary Table S2) to identify immune cell subsets according to standard techniques. For the staining of Supplementary Figure S7, heparinized blood of three healthy donors was isolated by Ficoll Paque™ Plus 206 (Amersham Pharmacia Biotech) density-gradient centrifugation. Peripheral blood mononuclear cells (PBMCs) were then stained with antibodies against CD19, CD24, CD27, CD38, IgM, IgG, IgA, and IgD (Supplementary Table S2). Cells were acquired on a BD FACSLyric™ (BD Biosciences). Data were analyzed with FlowJo ver. 10 (Treestar). Dead cells were excluded from analysis by side/forward scatter gating.
Carsetti R., Zaffina S., Piano Mortari E., Terreri S., Corrente F., Capponi C., Palomba P., Mirabella M., Cascioli S., Palange P., Cuccaro I., Milito C., Zumla A., Maeurer M., Camisa V., Vinci M.R., Santoro A., Cimini E., Marchioni L., Nicastri E., Palmieri F., Agrati C., Ippolito G., Porzio O., Concato C., Onetti Muda A., Raponi M., Quintarelli C., Quinti I, & Locatelli F. (2020). Different Innate and Adaptive Immune Responses to SARS-CoV-2 Infection of Asymptomatic, Mild, and Severe Cases. Frontiers in Immunology, 11, 610300.
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