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Ficoll paque

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Ficoll-Paque is a density gradient medium used for the isolation and purification of cells and cellular components. It is a sterile, pyrogen-free solution that allows for the separation of different cell types based on their density.

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Lab products found in correlation

Cryopure tube, by Sarstedt (1 mentions) Fetal bovine serum (fbs), by Harvard Bioscience (1 mentions) Dimethylsulfoxid dmso, by Merck Group (1 mentions) Ls column, by Miltenyi Biotec (1 mentions) Biocoll gradient, by Harvard Bioscience (1 mentions) Influx flow cytometry, by BD (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Ifn γ, by Immunotools (1 mentions) Gm csf, by Immunotools (1 mentions) Fetal calf serum (fcs), by Harvard Bioscience (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) 75 cm2 cell culture flasks, by BD (1 mentions) Interleukin 4 (il 4), by R&D Systems (1 mentions) Nocodazole, by Merck Group (1 mentions) Dimethyl sulfoxide (dmso), by ITW Reagents (1 mentions) Domatinostat, by ITW Reagents (1 mentions) Rpmi 1640 medium, by PAN Biotech (1 mentions) Vacutainer, by BD (1 mentions)

Spelling variants of this product

Ficoll (64 citations) Ficoll-Paque gradient centrifugation (3 citations) Ficoll-Paque density gradient (3 citations) Ficoll-Paque (Histopaque-1077; (2 citations)

18 protocols using ficoll paque

1

PBMC Isolation from Whole Blood

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Blood was collected in BD Vacutainer lithium heparin-treated tubes (BD Biosciences; San Jose, CA, USA), and diluted 1:1 with phosphate-buffered saline (PBS). Peripheral blood mononuclear cells (PBMCs) were prepared by density gradient centrifugation on Ficoll Paque (Biochrom AG, Berlin) at 375g for 20 min at room temperature. The cells were harvested and washed twice in PBS. Cells were then suspended in Fluorescence Assisted Cell Sorting (FACS) staining buffer (PBS w/0.5% BSA) and cell numbers determined.
Busse S., Steiner J., Glorius S., Dobrowolny H., Greiner-Bohl S., Mawrin C., Bommhardt U., Hartig R., Bogerts B, & Busse M. (2015). VGF expression by T lymphocytes in patients with Alzheimer's disease. Oncotarget, 6(17), 14843-14851.
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2

Cryopreservation of Kidney Transplant PBMCs

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Blood was collected from each kidney transplant recipient recruited into the study at least once and up to four times in Vacutainers containing lithium heparin (Beckton Dickinson (BD), Franklin Lakes, US-NJ). The PBMCs were isolated by density gradient centrifugation Ficoll-Paque (Biochrom, Berlin, Germany). The cells were frozen in CryoPure tubes (Sarstedt, Nümbrecht, Germany) in Roswell Parm Memorial Institute-1640 medium (RPMI, Biochrom) with 60% fetal bovine serum (FBS, Biochrom) and 10% dimethylsulfoxid (DMSO; Sigma-Aldrich, Munich, Germany) at −80 °C in a freezing box (ThermoFisher, Waltham, US-MA) containing isopropylalcohole (Sigma-Aldrich) and stored in liquid nitrogen until further usage. Upon defrosting, the cells were left resting for 24 hours.
Thieme C.J., Weist B.J., Mueskes A., Roch T., Stervbo U., Rosiewicz K., Wehler P., Stein M., Nickel P., Kurtz A., Lachmann N., Choi M., Schmueck-Henneresse M., Westhoff T.H., Reinke P, & Babel N. (2019). The TreaT-Assay: A Novel Urine-Derived Donor Kidney Cell-Based Assay for Prediction of Kidney Transplantation Outcome. Scientific Reports, 9, 19037.
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3

Peripheral Blood Sample Analysis for ALL

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Peripheral blood samples of ALL patients were obtained after written informed consent at the University of Tübingen. Peripheral blood mononuclear cells (PBMC) were isolated by Ficoll/Paque (Biochrom) density gradient centrifugation and viably stored in liquid nitrogen until analysis. This study was approved by the institutional review board to be in accordance with the ethical standards and the Declaration of Helsinki. Diagnosis of precursor B cell and T cell ALL was confirmed by morphologic analysis, immunophenotyping, and genetic features.
Rothfelder K., Hagelstein I., Roerden M., Blumenstock G., Hofmann M., Nuebling T., Jung G., Salih H.R, & Dörfel D. (2018). Expression of the Immune Checkpoint Modulator OX40 in Acute Lymphoblastic Leukemia Is Associated with BCR-ABL Positivity. Neoplasia (New York, N.Y.), 20(11), 1150-1160.
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4

Isolation of CD34+ Cells from Peripheral Blood

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CD34+ cells were isolated from low-density mononuclear cells (MNC) derived from peripheral blood of initiated healthy donors, using gradient isolation (1.077 g/ml Ficoll-Paque, Biochrom) and immunomagnetic selection, using a combination of the Miltenyi CD34 MultiSort kit (Miltenyi Biotec) and the LS Columns (Miltenyi Biotec) in accordance with the manufacturer’s instructions. In all preparations, CD34+ cell purity exceeded 98%, as estimated by flow cytometry (data not shown).
Stavrou E.F., Lazaris V.M., Giannakopoulos A., Papapetrou E., Spyridonidis A., Zoumbos N.C., Gkountis A, & Athanassiadou A. (2017). The β-globin Replicator greatly enhances the potential of S/MAR based episomal vectors for gene transfer into human haematopoietic progenitor cells. Scientific Reports, 7, 40673.
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5

Standardized PCC Assay Protocol

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Taking into consideration that the PCC assay is technically demanding and not yet widely used, a detailed protocol for PEG mediated cell fusion and PCC induction in lymphocytes isolated from the peripheral blood using mitotic CHO cells was established by the L1 laboratory and shared with the RENEB partners, in order to harmonize and standardize the PCC assay as well as the scoring criteria. Informed consent was obtained for each blood donor, and packaging, labelling and shipment of blood samples conformed to national and international regulations. A temperature logger, dosimeter to record the temperature and any dose received by the samples during transport, was used together with the standardized sample instruction sheet (ISO21243, 2008 , ISO19238, 2014 ). The peripheral blood was always sampled in heparinized vials and, for the isolation of lymphocytes, whole blood was carefully layered on top of equal amount of Ficoll-Paque or Biocoll gradient (Biochrom) in a test tube before centrifugation. In order to quantitate radiation exposure by means of initially induced PCC fragments, the PCC methodology was applied as soon as possible, since the initial radiation-induced fragments in excess of 46 PCCs decrease with time (Pantelias and Maillie, 1985a (link), Darroudi et al., 1998a (link)).
Terzoudi G.I., Pantelias G., Darroudi F., Barszczewska K., Buraczewska I., Depuydt J., Georgieva D., Hadjidekova V., Hatzi V.I., Karachristou I., Karakosta M., Meschini R., M’Kacher R., Montoro A., Palitti F., Pantelias A., Pepe G., Ricoul M., Sabatier L., Sebastià N., Sommer S., Vral A., Zafiropoulos D, & Wojcik A. (2016). Dose assessment inter-comparisons within the RENEB network using G0-lymphocyte prematurely condensed chromosomes (PCC assay). International journal of radiation biology, 93(1), 48-57.
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6

HEATR1 expression in glioblastoma

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A total of 22 frozen GBM tumor tissues were obtained from the Department of Neurosurgery, Huashan Hospital, to analyze the expression level of HEATR1 mRNA. Additionally, eight control brain tissue samples were obtained from adjacent brain tissues of patients with traumatic brain injury who suffered contusion and laceration. In addition, 10 GBM formalin-fixed, paraffin-embedded (FFPE) tissue sections and 10 normal brain tissues were analyzed by IHC.
Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll/Paque (Biochrom, Berlin, Germany) density gradient centrifugation of heparinized blood obtained from healthy donors (n = 6) and patients (benign tumors, 5; grade 2 astrocytoma, 7; grade 3 anaplastic glioma, 10; glioblastoma, 16). The patients' clinical characteristics are listed in Table 1.
Wu Z.B., Qiu C., Zhang A.L., Cai L., Lin S.J., Yao Y., Tang Q.S., Xu M., Hua W., Chu Y.W., Mao Y., Zhu J.H., Xu J, & Zhou L.F. (2014). Glioma-Associated Antigen HEATR1 Induces Functional Cytotoxic T Lymphocytes in Patients with Glioma. Journal of Immunology Research, 2014, 131494.
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7

Isolation and Culture of Human CD4+ T Cells

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Human peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood of healthy patients, using Ficoll‐Paque (Biochrom, Berlin, Germany) density gradient centrifugation. Then PBMCs incubated with CD4‐APC antibody (BD Biosciences, CA) for 20 minutes. Human CD4+ T cells were isolated from PBMCs via BD influx flow cytometry (BD Biosciences). CD4+ T cells were cultured in RPMI‐1640 containing 10% FBS supplemented with antibiotics.
Xiang P., Jin S., Yang Y., Sheng J., He Q., Song Y., Yu W., Hu S, & Jin J. (2019). Infiltrating CD4+ T cells attenuate chemotherapy sensitivity in prostate cancer via CCL5 signaling. The Prostate, 79(9), 1018-1031.
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8

Peripheral Blood Sampling and Analysis

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Peripheral blood samples (0.5–7 mL) were collected in EDTA-coated tubes. The concentration of leukocytes, their populations, and platelets was determined using a hematology analyzer (Cell-Dyn 3700, Abbot Pharmaceutical Co. Ltd., Lake Bluff, IL, USA). PBMCs were isolated from whole blood by centrifugation over a Ficoll-Paque gradient (Biochrom Ltd., Cambridge, UK) and counted in a hematology analyzer (Cell-Dyn 3700). Plasma was collected and stored at −80°C.
Thomas I., Panagoulias I., Aggeletopoulou I., Varvarigou A., Spiliotis B.E, & Mouzaki A. (2021). The Role of Leptin in Childhood Immune Thrombocytopenia (ITP): An Anti-Inflammatory Agent?. International Journal of Molecular Sciences, 22(14), 7636.
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9

Isolation and Differentiation of Human Macrophages

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Peripheral blood mononuclear cells (PBMC) were isolated from buffy coats of healthy donors by Ficoll-Paque (Biochrom) density centrifugation. The local ethics committee of the University of Bonn approved the human PBMC study protocol. Monocytes were enriched using the CD14 kit (Miltenyi Biotec). A purity of more than 98% was achieved, as assessed by flow cytometry. Monocytes were cultured at a density of 1.4 × 106/mL in RPMI1640 (Gibco) supplemented with 10% FCS for 5 days. The lineage-determining GMCSF (50 U/mL or 20 ng/mL; ImmunoTools) was added for macrophage maturation. Cells were either left as untreated (monocyte-derived macrophages) or the following additional stimuli were added: (i) TAMs were induced by a 5-day incubation with cell-free supernatant from the VSCC cell cultures (40%, in vitro polarized TAMs); and (ii) M1 macrophages were induced by adding 20 ng/mL IFNγ (ImmunoTools) for 5 days.
Condic M., Rohr A., Riemann S., Staerk C., Ayub T.H., Doeser A., Zillinger T., Merkelbach-Bruse S., Buettner R., Barchet W., Rudlowski C., Mustea A, & Kübler K. (2024). Immune Profiling of Vulvar Squamous Cell Cancer Discovers a Macrophage-rich Subtype Associated with Poor Prognosis. Cancer Research Communications, 4(3), 861-875.
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10

HLA-A*02:01-specific T-cell Isolation

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The clinical material was used with approval by Charité ethics committee (Approval No. EA1/222/14 and EA1/026/14) and written informed consent by the patients and volunteer donors. All methods and procedures were performed in accordance with the relevant guidelines and regulations including the handling of primary clinical materials, and anonymization and processing of patient-related information and data. HLA typing was carried out by the HLA typing laboratory, Charité - Universitätsmedizin Berlin. Peripheral blood of 4 HLA-A*02:01-positive healthy volunteers and 8 HLA-A*02:01-positive chronic myeloid leukemia (CML) patients was used to isolate PBMCs by density centrifugation using Ficoll Paque (Biochrom, Berlin, Germany) and cryopreserved in FCS (Biochrom, Berlin, Germany) with 10% DMSO (Pierce, Illinois, USA) at −140 °C. For in vitro priming, PBMCs were pulsed with 10 µg/ml of each peptide in ExVivo 15 serum-free medium (Biowhitaker, Belgium) at 37 °C and 8% CO2. 50 U/ml of recombinant human IL-2 (Chiron, Munich, Germany) was added to the cultures on day 1 and day 3. On day 8 the primed PBMCs were harvested, washed with PBS (Gibco, Grand Island, NY, USA) and counted before analysis in INFγ ELISpot assays detailed in supplementary methods and by flow cytometry.
Nyambura L.W., Muñoz A.A., le Coutre P, & Walden P. (2019). HLA class I-restricted T cell epitopes isolated and identified from myeloid leukemia cells. Scientific Reports, 9, 14029.
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