Ficoll hypaque

Manufactured by GE Healthcare

Sourced in United States, Sweden, United Kingdom, Germany, China, Norway, Spain, Canada, Italy, Austria

Ficoll-Hypaque is a density gradient medium used for the isolation of mononuclear cells from whole blood, bone marrow, or other tissues. It is a sterile, pyrogen-free solution consisting of Ficoll polymer and sodium diatrizoate.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Ficoll (495 citations) Ficoll-Hypaque density gradient centrifugation (34 citations) Ficoll-Hypaque density gradient (27 citations) Ficoll-Hypaque solution (23 citations) Ficoll-Hypaque density (9 citations) Ficoll-Hypaque PREMIUM (8 citations) Ficoll-Hypaque gradient centrifugation (7 citations) Ficoll-Hypaque cushion (6 citations) Ficoll-Hypaque plus density gradient (3 citations) Ficoll-Hypaque Plus gradient (3 citations)

553 protocols using ficoll hypaque

Isolation of Lamina Propria Leukocytes and Peripheral Blood Mononuclear Cells

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

Lamina propria leukocytes (LPL) were isolated by enzymatic tissue digestion according to a modified method of Bull and Bookman [14] (link). Briefly, after depletion of the epithelial layer (see above) mucosal tissue was cut into 2–4 mm pieces and digested in a shaking waterbath at 37°C for 1.5 h using collagenase IV (70 µg/ml; Sigma) and deoxyribonuclease I (100 µg/ml; Sigma) in RPMI 1640 containing 2% fetal calf serum (Sigma), 2% L-glutamine (Life Technologies), and antibiotics. The resulting cell suspension was separated from undigested tissue by filtration through a 70 µm nylon mesh (BD Bioscience, Heidelberg, Germany). For further purification, the cell suspension was subjected to Percoll (GE Healthcare, Munich, Germany) and Ficoll-Hypaque (GE Healthcare) density gradient centrifugation.
Peripheral blood was taken during the operation. Peripheral blood mononuclear cells (PBL) were obtained by Ficoll-Hypaque (GE Healthcare) density gradient centrifugation.

Schröder-Braunstein J., Gras J., Brors B., Schwarz S., Szikszai T., Lasitschka F., Wabnitz G., Heidtmann A., Lee Y.S., Schiessling S., Leowardi C., Al-Saeedi M., Ulrich A., Engelke A., Winter J., Samstag Y., Giese T, & Meuer S. (2014). Initiation of an Inflammatory Response in Resident Intestinal Lamina Propria Cells -Use of a Human Organ Culture Model. PLoS ONE, 9(5), e97780.

+ Open protocol

+ Expand

Isolation and Culture of Endothelial Progenitor Cells

Cited 1 time

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

A human peripheral blood sample was collected from a healthy donor^{14 (link)}. Peripheral blood mononuclear cells (PBMCs) were separated by Ficoll-Hypaque (GE Healthcare Life Science, Piscataway, NJ, USA) density gradient centrifugation at 2500 rpm for 30 min, and were obtained from the interface between the plasma layer and Ficoll-Hypaque layer^{3 (link),15}. PBMCs were seeded into a fibronectin-coated T25 flask at 1–3 × 10⁷ cells with Defined Keratinocyte-SFM (Gibco, Waltham, MA, USA)-based medium containing 0.2 mM ascorbic acid, 10 µg/ml l-glutamine, 10 ng/ml human epidermal growth factor, 5 µg/ml insulin, 1 ng/ml selenium, 74 ng/ml hydrocortisone, 5 ng/ml Lin28, 1% antibiotic-antimycotic, and 10% fetal bovine serum were incubated at 37°C, 5% CO₂. Medium was changed on day 2 (day 0; the day when EPCs were seeded) and then replaced twice a week. EPC colony formation appeared after 2–4 weeks incubation. EPC colonies were passaged to T25 flasks or 6-well plates according to colony size. Isolated EPCs were passaged when they reached 70–90% confluence.

Lee S.H., Ra J.C., Oh H.J., Kim M.J., Setyawan E.M., Choi Y.B., Yang J.W., Kang S.K., Han S.H., Kim G.A, & Lee B.C. (2019). Clinical Assessment of Intravenous Endothelial Progenitor Cell Transplantation in Dogs. Cell Transplantation, 28(7), 943-954.

+ Open protocol

+ Expand

Isolation of Human Peripheral Blood Mononuclear Cells

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

Human specimens were collected with written consent from volunteers in accordance with institutional review board guidelines and approval. Phlebotomy was conducted on healthy donors; 6–12 ml of peripheral blood were withdrawn into 6 ml tubes (BD Vacutainer® EDTA, purple cap) and spun down for 10 min at 1300 g at room temperature (RT) without centrifuge brake, this allowed plasma separation. The non-plasma fraction was diluted 1:1 with RPMI and carefully laid on top of Ficoll-Hypaque (GE Healthcare) solution (1 ml of Ficoll-Hypaque per 3 ml of blood/RPMI mixture) in separate 15 ml-tubes, and centrifugated at 800 g for 30 min at RT without brake in swinging-bucket rotor. The mononuclear cell layer was carefully extracted and washed 3 times with RPMI by 300 g centrifugation for 5 min at 4 °C. PBMCs were counted and used fresh for co-culture experiments or frozen by resuspension at 5 × 10⁶ cells/ml in 90% FBS/10% DMSO.

Vicencio J.M., Evans R., Green R., An Z., Deng J., Treacy C., Mustapha R., Monypenny J., Costoya C., Lawler K., Ng K., De-Souza K., Coban O., Gomez V., Clancy J., Chen S.H., Chalk A., Wong F., Gordon P., Savage C., Gomes C., Pan T., Alfano G., Dolcetti L., Chan J.N., Flores-Borja F., Barber P.R., Weitsman G., Sosnowska D., Capone E., Iacobelli S., Hochhauser D., Hartley J.A., Parsons M., Arnold J.N., Ameer-Beg S., Quezada S.A., Yarden Y., Sala G, & Ng T. (2022). Osimertinib and anti-HER3 combination therapy engages immune dependent tumor toxicity via STING activation in trans. Cell Death & Disease, 13(3), 274.

+ Open protocol

+ Expand

Isolation of CD3+ Lymphocytes from Patient Blood

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 the peripheral blood of patients prior to lymphodepletion by density gradient with Ficoll-Hypaque (Pharmacia Biotech, Piscataway, NJ, USA). CD3⁺ untouched lymphocytes were separated from the peripheral blood of patients before lymphodepletion by RosetteSep^TM Human T Cell Enrichment Cocktail (StemCell Technologies, Vancouver, Canada) using negative selection. Briefly, 4 mL of whole blood was mixed with 200 µL RosetteSep™ Cocktail and centrifuged over a density gradient medium (Ficoll-Hypaque, Pharmacia Biotech, Piscataway, NJ, USA). CD3⁺ enriched population at the interface between the plasma and the density gradient medium was collected.

Beider K., Itzhaki O., Schachter J., Grushchenko-Polaq A.H., Voevoda-Dimenshtein V., Rosenberg E., Ostrovsky O., Devillers O., Shapira Frommer R., Zeltzer L.A., Toren A., Jacoby E., Shimoni A., Avigdor A., Nagler A, & Besser M.J. (2022). Molecular and Functional Signatures Associated with CAR T Cell Exhaustion and Impaired Clinical Response in Patients with B Cell Malignancies. Cells, 11(7), 1140.

+ Open protocol

+ Expand

Lymphocyte Isolation and Formalin Fixation

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

Blood samples (5 mL) were collected in buffered EDTA, diluted with an equal volume of phosphate buffered saline (PBS) solution (Welgene, Daegu, Korea), and layered onto Ficoll-Hypaque (Ficoll-Paque, GE Healthcare Bio-Sciences, Pittsburgh, PA). The samples were centrifuged in 15 mL disposable centrifuge tubes at 2,000 rpm for 20 minutes. After drawing off the upper layer, the lymphocyte layer was transferred to a 50 mL centrifuge tube, to which was added 3 volumes of PBS followed by centrifugation at 1,500 rpm for 10 minutes. After removing the supernatant, the cell pellet was mixed with an equal volume of 30 mg/mL gelatin (Sigma-Aldrich, Seoul, Korea) and the mixture (gel block) was refrigerated for 20 minutes at 4ºC and fixed with neutralized buffered formalin overnight. The fixed cell pellet mixture was wrapped in crayon paper, placed in a cassette, and stored in 80% ethanol. The pellets were next processed in the automatic tissue processor using a 13-hour schedule, consisting of 80% ethanol with 1 change (2.5 hours), 95% ethanol (1 hour), 100% ethanol, with four changes (1 hour each), 1:1 ethanol/xylene (1 hour); xylene with three changes (1 hour each), paraffin wax at 60°C (1 hour); and paraffin wax at 60°C with vacuum impregnation at 20 lb (30 minutes).

Nam S.J., Yeo H.Y., Chang H.J., Kim B.H., Hong E.K, & Park J.W. (2016). A New Cell Block Method for Multiple Immunohistochemical Analysis of Circulating Tumor Cells in Patients with Liver Cancer. Cancer Research and Treatment : Official Journal of Korean Cancer Association, 48(4), 1229-1242.

+ Open protocol

+ Expand

Isolation and Polarization of M2 Macrophages

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

Fresh blood samples from healthy donors were collected, and PBMCs were isolated by density gradient centrifugation using Ficoll‐Hypaque (GE Healthcare, NJ, USA). Isolation of monocytes and induction into M0 macrophages were done using the above procedures. Next, human recombinant IL‐33 (PeproTech, New Jersey, USA) was used to induce M0 macrophages to differentiate into M2 macrophages for 24 hours. All cells were cultured at 37°C in a humidified incubator with 5% CO₂.

Mai S., Liu L., Jiang J., Ren P., Diao D., Wang H, & Cai K. (2020). Oesophageal squamous cell carcinoma–associated IL‐33 rewires macrophage polarization towards M2 via activating ornithine decarboxylase. Cell Proliferation, 54(2), e12960.

+ Open protocol

+ Expand

Isolation and Differentiation of Human CD14+ Monocytes

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

This experiment was approved by the Institutional Review Board of Kaohsiung Medical University Hospital (KMUH-IRB-20140303). CD14⁺ monocytes were collected from human peripheral blood mononuclear cells (PBMCs) obtained from blood buffy coats according to the previously described protocol [29 (link),30 (link)]. Blood sample (20 mL) from the veins of healthy volunteers was collected into sterile tubes containing EDTA. According to the manufacturer’s instructions, PBMCs were separated by gradient centrifugation on Ficoll–Hypaque (GE Healthcare, Wauwatosa, WI, USA) containing anti-CD14 microbeads (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany). Cells were washed twice with phosphate buffered saline (PBS) and then resuspended in Roswell Park Memorial Institute (RPMI) 1640 medium (Mediatech, Manassas, VA, USA) containing 10% FBS, 100 μg/mL streptomycin, 100 U/mL penicillin, and 0.25 g/mL Amphotericin B. Cells were labeled with CD3-PE and CD14-FITC and then assessed for the purity of CD14⁺ cells by flow cytometry, which showed >95% purity. The purified human CD14⁺ cells were differentiated into macrophages by adding 10 ng/mL human granulocyte-macrophage colony-stimulating factor (GM-CSF) (PeproTech, Rocky Hill, NJ, USA) for 6 days at 37 °C in 5% CO₂, and were prepared for subsequent experiments.

Lin C.Y., Kao S.H., Hung L.C., Chien H.J., Wang W.H., Chang Y.W, & Chen Y.H. (2021). Lipopolysaccharide-Induced Nitric Oxide and Prostaglandin E2 Production Is Inhibited by Tellimagrandin II in Mouse and Human Macrophages. Life, 11(5), 411.

+ Open protocol

+ Expand

Isolation of immune cell subsets

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

T cells, B cells, NK cells and monocytes for NFAM1 expression experiments were isolated from human peripheral blood that was obtained from the Boehringer Ingelheim blood donor program. Using the standardized Ficoll-Hypaque (GE Healthcare) density gradient centrifugation method, PBMCs were isolated from whole blood. PBMCs were then frozen at -80 degrees Celsius until further use. T cells, B cells, NK cells and monocytes were isolated from thawed PBMCs via the use of Stemcell Technologies enrichment kits (#19051, #19054, #19055 and #19058, respectively). Neutrophils were isolated directly from human whole blood via the use of Stemcell Technologies neutrophil enrichment kit (#19257, RBC lysis method).

Juchem K.W., Gounder A.P., Gao J.P., Seccareccia E., Yeddula N., Huffmaster N.J., Côté-Martin A., Fogal S.E., Souza D., Wang S.S., Glynn E.R., Yung I., Ritchie J., Li L., Zheng J., Mbow M.L., Li J, & Chanda S.K. (2022). NFAM1 Promotes Pro-Inflammatory Cytokine Production in Mouse and Human Monocytes. Frontiers in Immunology, 12, 773445.

+ Open protocol

+ Expand

Isolation and Cryopreservation of PBMCs

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

Peripheral blood mononuclear cells (PBMCs) were isolated by the Ficoll-Hypaque density gradient centrifugation (GE Healthcare) of venous blood samples obtained from healthy volunteers, the patient, his parents, and his six-year-old brother. Cells were cryopreserved and stored at −150°C until use.

Ogishi M., Yang R., Aytekin C., Langlais D., Bourgey M., Khan T., Ali F.A., Rahman M., Delmonte O.M., Chrabieh M., Zhang P., Gruber C., Pelham S.J., Spaan A.N., Rosain J., Lei W.T., Drutman S., Hellmann M.D., Callahan M.K., Adamow M., Wong P., Wolchok J.D., Rao G., Ma C.S., Nakajima Y., Yaguchi T., Chamoto K., Williams S.C., Emile J.F., Rozenberg F., Glickman M.S., Rapaport F., Kerner G., Allington G., Tezcan I., Cagdas D., Hosnut F.O., Dogu F., Ikinciogullari A., Rao V.K., Kainulainen L., Béziat V., Bustamante J., Vilarinho S., Lifton R.P., Boisson B., Abel L., Bogunovic D., Marr N., Notarangelo L.D., Tangye S.G., Honjo T., Gros P., Boisson-Dupuis S, & Casanova J.L. (2021). Inherited PD-1 deficiency underlies tuberculosis and autoimmunity in a child. Nature medicine, 27(9), 1646-1654.

+ Open protocol

+ Expand

CD4+ T Cell Activation and Isolation

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

Human PBMCs were isolated using Ficoll-Hypaque (GE-Healthcare; GE17–1440-02) gradient separation. For RNA-seq CD4⁺ T cells were enriched using CD4 microbeads (Miltenyi; 130–045-101) and 4×10⁶ cells/ml were resuspended in RPMIc with 50 U/ml IL-2 (Immunotools; 11340023) in 24-well plate wells and stimulated for 20h with 25µl/ml ImmunoCult CD3/CD28 T cell activator (Stemcell; 10971) prior to staining and cell sorting.

Klicznik M.M., Morawski P.A., Höllbacher B., Varkhande S.R., Motley S., Kuri-Cervantes L., Goodwin E., Rosenblum M.D., Long S.A., Brachtl G., Duhen T., Betts M.R., Campbell D.J, & Gratz I.K. (2019). Human CD4+CD103+ cutaneous resident memory T cells are found in the circulation of healthy subjects. Science immunology, 4(37), eaav8995.

+ 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!