Ficoll density centrifugation

Manufactured by GE Healthcare

Sourced in New Zealand, Germany, Sweden, United States

Ficoll density centrifugation is a laboratory technique used to separate different types of cells or particles based on their densities. It involves the use of a Ficoll solution, a synthetic polymer, to create a density gradient. During centrifugation, the different components in the sample will migrate to the layers of the gradient that match their respective densities, allowing for their separation and isolation.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Ficoll density gradient centrifugation kit Ficoll-Hypaque density gradient centrifugation Ficoll-Paque PLUS density gradient centrifugation Ficoll isopaque density gradient centrifugation Ficoll-Paque Plus density centrifugation Ficoll plus density gradient centrifugation Ficoll-Hypaque density centrifugation Ficoll density gradient centrifugation Ficoll-Paque density gradient centrifugation Ficoll-Paque density centrifugation

19 protocols using ficoll density centrifugation

Isolation of DLBCL and Healthy B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Frozen DLBCL patient tumor biopsy samples were obtained according to guidelines approved by the Institutional Review Board of the Cleveland Clinic. DLBCL patients were identified as GCB or ABC based on application of the Hans immunochemistry algorithm to fresh-frozen paraffin-embedded tumor biopsies.^{31 (link)} B cells were purified from PBMCs of healthy individuals using the human B cell isolation kit (Miltenyi Biotec). Tonsils were obtained from healthy individuals undergoing routine tonsillectomies in accordance with ethical recommendations. After mincing, tonsillar mononuclear cells were isolated by Ficoll density centrifugation (GE Healthcare). GC B cells were enriched by magnetic cell separation with anti-CD77, MARM-4 (Abcam) and IgG1 microbeads (Miltenyi Biotec).

Pore D., Bodo J., Danda A., Yan D., Phillips J.G., Lindner D., Hill B.T., Smith M.R., Hsi E.D, & Gupta N. (2015). Identification of Ezrin-Radixin-Moesin proteins as novel regulators of pathogenic B cell receptor signaling and tumor growth in diffuse large B cell lymphoma. Leukemia, 29(9), 1857-1867.

+ Open protocol

+ Expand

Fibroblast Culture and SOCE-deficient PBMC

Check if the same lab product or an alternative is used in the 5 most similar protocols

Patient and healthy donor (HD) fibroblasts were cultured in RPMI 1640 medium supplemented with 10% FCS, 1% L-glutamine, 1% HEPES, and 1% Penicillin/Streptomycin (all from Mediatech, Manassas, VA). SOCE-deficient PBMC from P3 and P6 before their first and second HSCT, respectively, and a HD were isolated from whole blood by Ficoll density centrifugation (GE Healthcare, Marlborough, MA) and stimulated with irradiated buffy coat cells, irradiated B cells, PHA (1 μg/ml) and recombinant human IL-2 (50 U/ml) for 10–12 days.

Lian J., Cuk M., Kahlfuss S., Kozhaya L., Vaeth M., Rieux-Laucat F., Picard C., Benson M.J., Jakovcevic A., Bilic K., Martinac I., Stathopulos P., Kacskovics I., Vraetz T., Speckmann C., Ehl S., Issekutz T., Unutmaz D, & Feske S. (2017). ORAI1 mutations abolishing store-operated Ca2+ entry cause anhidrotic ectodermal dysplasia with immunodeficiency (EDA-ID). The Journal of allergy and clinical immunology, 142(4), 1297-1310.e11.

+ Open protocol

+ Expand

Isolation of NK and T Cells from Human PBMCs

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human peripheral blood mononuclear cells (PBMCs) were freshly prepared by Ficoll density centrifugation (GE Health) from blood that was collected from 11 health donors as described previously with some modifications [17 (link),43 (link)]. According to the manufacturer's instructions, NK cells or T cells were then isolated from the PBMCs by EasySepTM Human NK Cell Enrichment Kit (Stem cell Co. Ltd, cat# 1905) or EasySepTM Human T Cell Enrichment Kit (Stem cell Co. Ltd, cat#19,661).

Li Y., Wu L., Liu Y., Ma S., Huang B., Feng X, & Wang H. (2022). A novel multifunctional anti-PD-L1-CD16a-IL15 induces potent cancer cell killing in PD-L1-positive tumour cells. Translational Oncology, 21, 101424.

+ Open protocol

+ Expand

Standardized PBMC Isolation and Cryopreservation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Complete blood counts including TLC were serially performed by the certified clinical laboratory of the recruiting hospital for each patient. PBMC were isolated from whole blood (Ficoll density centrifugation; GE Healthcare) and cryopreserved using rigorous standardized operating procedures (SOPs) developed and validated by our Experimental Therapeutics Program for all steps of sample procurement, processing, cryopreservation, storage and subsequent thawing. Following our strict SOPs for PBMC isolation and cryopreservation^{36 (link)}, the majority of thawed samples had viabilities of over 90%. Any sample with cell viability of less than 70% was excluded from analysis.

Ghadiri M., Rezk A., Li R., Evans A., Giacomini P.S., Barnett M.H., Antel J, & Bar-Or A. (2020). Pre-treatment T-cell subsets associate with fingolimod treatment responsiveness in multiple sclerosis. Scientific Reports, 10, 356.

+ Open protocol

+ Expand

Isolation of DLBCL and Healthy B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

Isolation and Culture of CD4+ T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mononuclear cells (PBMCs) were isolated from peripheral blood by Ficoll density centrifugation (Cat# 45-001-750, GE Healthcare, Piscataway, NJ). CD4 T cells were isolated from PBMCs using the CD4 T cell negative selection kit (Cat# 130-096-533, Miltenyi Biotec, Auburn, CA). The cells were cultured in RPMI-1640 medium supplemented with 10% FBS (Cat# S11050H, Atlanta Biologicals, Flowery Branch, GA), 100 IU/ml penicillin, and 2 mM L-glutamine (Cat# 25-030-081, Thermo Fisher Scientific, Waltham, MA) and maintained at 37°C in 5% CO₂ incubator.

Dang X., Cao D., Zhao J., Schank M., Khanal S., Nguyen L.N., Wu X.Y., Zhang Y., Zhang J., Jiang Y., Ning S., Wang L., El Gazzar M., Moorman J.P, & Yao Z.Q. (2022). Mitochondrial topoisomerase 1 inhibition induces topological DNA damage and T cell dysfunction in patients with chronic viral infection. Frontiers in Cellular and Infection Microbiology, 12, 1026293.

+ Open protocol

+ Expand

PBMC Isolation and Viability Assessment

Check if the same lab product or an alternative is used in the 5 most similar protocols

PBMCs were isolated by Ficoll density centrifugation (GE Healthcare, Solingen, Germany) from EDTA blood samples drawn from the dialysis fistula before the dialysis session. The quality of isolated cells was tested by 7-AAD staining Thermo Fisher Scientific, Darmstadt, Germany). The vitality of PBMC was 99.2% ± 1.6 for N and 99.1% ± 0.5 for H patients.

Ulrich C., Fiedler R., Herberger E., Canim Z., Markau S, & Girndt M. (2024). Hypervolemia in Dialysis Patients Impairs STAT3 Signaling and Upregulates miR-142-3p: Effects on IL-10 and IL-6. International Journal of Molecular Sciences, 25(7), 3719.

+ Open protocol

+ Expand

Regulation of T Cell Apoptosis and DNA Damage

Check if the same lab product or an alternative is used in the 5 most similar protocols

Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood by Ficoll density centrifugation (GE Healthcare, Piscataway, NJ). CD4⁺ T cells were isolated from PBMCs using the CD4⁺ T Cell Negative Selection Kit (Miltenyi Biotec Inc., Auburn, CA). T cells were cultured in RPMI 1640 medium containing 10% FBS (Atlanta Biologicals, Flowery Branch, GA), 100 IU/ml penicillin, and 2 mM L-glutamine (Thermo Scientific, Logan, Utah) at 37°C and 5% CO₂ atmosphere. Cells were harvested after day 1 or day 4 of culture for the detection of apoptosis and DNA damage. To test the role of ATM activation in repairing DNA damage and apoptosis, 10 μM ATM inhibitor (KU60019, Abcam, Cambridge, MA) or DMSO were added to the cultures at day 1, and the cells were collected after 48 h for measuring apoptosis and DNA damage. To determine the role of the PI3K pathway in ATM activation, purified CD4 T cells were cultured for 48 h in the presence or absence of 20 μM PI3K inhibitor (LY294002, Sigma), followed by flow cytometry or western blot analysis.

Zhao J., Nguyen L.N., Nguyen L.N., Dang X., Cao D., Khanal S., Schank M., Thakuri B.K., Ogbu S.C., Morrison Z.D., Wu X.Y., Li Z., Zou Y., El Gazzar M., Ning S., Wang L., Moorman J.P, & Yao Z.Q. (2019). ATM Deficiency Accelerates DNA Damage, Telomere Erosion, and Premature T Cell Aging in HIV-Infected Individuals on Antiretroviral Therapy. Frontiers in Immunology, 10, 2531.

+ Open protocol

+ Expand

Immune Profiling in Parkinson's Disease

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antecubital venous blood was obtained contemporaneously from neurologically normal controls and participants with PD and used for fresh whole blood analysis and subsequent analysis of peripheral blood mononuclear cells (PBMCs) that were isolated by Ficoll density centrifugation (GE Healthcare) and then cryopreserved. All samples were processed at the University of Pennsylvania, with samples from the Columbia University samples (both NC and PD) overnight shipped to the University of Pennsylvania for next-day analysis and processing, using Credo Box shippers (Pelican BioThermal) that ensure temperature stability during transport. All steps of sample handling, shipping, processing and storage, and whole blood staining and PBMC assays were performed using the identical standard operating procedures (SOPs). Although the fresh whole blood analysis enables assessment of immune cell types that are less amenable to freeze and thaw, the cryopreserved PBMCs were thawed and cultured in batch, with samples selected to ensure age and sex balance between patients with PD and NCs, serving to both limit interassay variability and minimize batch effects.^{20 (link)}

Li R., Tropea T.F., Baratta L.R., Zuroff L., Diaz-Ortiz M.E., Zhang B., Shinoda K., Rezk A., Alcalay R.N., Chen-Plotkin A, & Bar-Or A. (2021). Abnormal B-Cell and Tfh-Cell Profiles in Patients With Parkinson Disease: A Cross-sectional Study. Neurology® Neuroimmunology & Neuroinflammation, 9(2), e1125.

+ Open protocol

+ Expand

Immune Profiling of COVID-19 Recovered Patients

Check if the same lab product or an alternative is used in the 5 most similar protocols

The study subjects were composed of two groups: 11 mild to moderate, non-hospitalized COVID-19-recovered patients and 4 control subjects, including 2 healthy subjects (HS) and 2 Influenza (Flu) patients. All COVID-19 patients were diagnosed using a positive PCR nucleic acid test and had recovered at least 2 weeks after the diagnosis. Blood from HS was obtained from BioIVT (Gray, TN) and was free of HBV, HCV, and HIV infections. The healthy control samples and Flu samples were confirmed by a negative serology test. PBMCs were isolated from 30-ml of blood by Ficoll density centrifugation (GE Healthcare; Piscataway, NJ) and stored in liquid nitrogen until used. The characteristics of the COVID-19 subjects are shown in Table 1.Table 1

Characteristics of COVID-19 patients.

Table 1

ID	Age	Gender	Symptom (Self-assessment)	Hospitalized (Y/N)	Sampling (Day)	Past Medical History
P1	54	F	Mild	N	74	N/A
P2	54	M	Mild	N	72	N/A
P3	51	F	Mild	N	78	N/A
P4	47	F	Moderate	N	75	Asthma
P5	20	F	Mild	N	77	N/A
P6	30	M	Moderate	N	88	N/A
P7	46	F	Moderate	N	119	Cardiomyopathy
P8	33	M	Mild	N	24	Hypertension, HIV
P9	42	F	Moderate	N	29	N/A
P10	42	M	Moderate	N	32	N/A
P16	47	M	Mild	N	15	Hypertension

Zhao J., Wang L., Schank M., Dang X., Lu Z., Cao D., Khanal S., Nguyen L.N., Nguyen L.N., Zhang J., Zhang Y., Adkins J.L., Baird E.M., Wu X.Y., Ning S., Gazzar M.E., Moorman J.P, & Yao Z.Q. (2021). SARS-CoV-2 specific memory T cell epitopes identified in COVID-19-recovered subjects. Virus Research, 304, 198508.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!