Ficoll paque density gradient

Manufactured by STEMCELL

Ficoll-Paque density gradient is a sterile, pyrogen-free colloidal polysaccharide solution used for the isolation of mononuclear cells from whole blood or bone marrow. It creates a density gradient that allows the separation of different cell types based on their density.

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Lab products found in correlation

Easysep human cd34 positive selection kit 2, by STEMCELL (3 mentions) Sepmate isolation tubes, by STEMCELL (1 mentions) Ficoll paque premium density gradient medium, by GE Healthcare (1 mentions)

5 protocols using ficoll paque density gradient

CD34+ Cell Isolation from Cord Blood

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Cord blood (CB) samples and AML patient samples were obtained with informed consent under local ethical approval, research ethics committee 07MRE05-44 (Cambridge) and ethics committee code number 94/2016/O/Tess (Bologna). Mononuclear cells (MNC) were isolated by Ficoll-Paque density gradient (StemCell Technologies) centrifugation. CB MNC were enriched for CD34⁺ cells using the RosetteSep Human CB CD34 Pre-Enrichment Cocktail (Stem Cell Technologies) as per manufacturer’s instructions.

Arede L., Foerner E., Wind S., Kulkarni R., Domingues A.F., Giotopoulos G., Kleinwaechter S., Mollenhauer-Starkl M., Davison H., Chandru A., Asby R., Samarista R., Gupta S., Forte D., Curti A., Scheer E., Huntly B.J., Tora L, & Pina C. (2022). KAT2A complexes ATAC and SAGA play unique roles in cell maintenance and identity in hematopoiesis and leukemia. Blood Advances, 6(1), 165-180.

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Isolation and Purification of CD34+ HSPCs

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CD34⁺ HSPCs from various stages of human development were isolated by positive magnetic selection using the EasySep Human CD34 Positive Selection Kit II (Stem Cell Technologies) after mononuclear cell isolation on a Ficoll-Paque density gradient (Stem Cell Technologies). Purity of isolated cells was assessed by flow cytometry with a PE conjugated anti-human CD34 antibody (Supplementary Table 3). Freshly isolated CD34⁺ HSPCs were either immediately cultured or cryopreserved for later use.

Basak A., Munschauer M., Lareau C.A., Montbleau K.E., Ulirsch J.C., Hartigan C.R., Schenone M., Lian J., Wang Y., Huang Y., Wu X., Gehrke L., Rice C.M., An X., Christou H.A., Mohandas N., Carr S.A., Chen J.J., Orkin S.H., Lander E.S, & Sankaran V.G. (2020). Control of human hemoglobin switching by LIN28B-mediated regulation of BCL11A translation. Nature genetics, 52(2), 138-145.

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Isolation and Cryopreservation of CD34+ HSPCs

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CD34⁺ HSPCs from human cord blood were isolated by positive magnetic selection using the EasySep Human CD34 Positive Selection Kit II (Stem Cell Technologies) after mononuclear cell isolation on a Ficoll-Paque density gradient (Stem Cell Technologies). The purity of isolated cells was assessed by flow cytometry with a conjugated anti-human CD34 antibody. Freshly isolated CD34⁺ HSPCs were either immediately cultured or cryopreserved for later use.

Shen Y., Li R., Teichert K., Montbleau K.E., Verboon J.M., Voit R.A, & Sankaran V.G. (2021). Pathogenic BCL11A variants provide insights into the mechanisms of human fetal hemoglobin silencing. PLoS Genetics, 17(10), e1009835.

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Rabbit and Bovine PBMC Isolation

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Rabbit PBMCs were isolated from 5 mL blood collected from the central artery of the rabbit ear before infection and at various time-points post-infection. Immediately after euthanasia, single-cell suspensions were prepared from popliteal lymph nodes and spleen as follows. Tissues were gently disrupted with scissors in sterile RPMI-1640 medium and passed through a 70 μm cell-strainer using a 1-mL syringe plunger. Mononuclear leukocyte suspensions were prepared from peripheral blood and tissue samples using Ficoll-Paque Premium density gradient medium (GE Healthcare). Each 5 mL single-cell suspension was diluted 1:1 in sterile PBS, overlaid onto a 5 mL Ficoll-Paque density cushion, and centrifuged (1825×g) for 20 min at room temperature. Mononuclear leukocytes at the interface were collected and washed in ice-cold PBS before further analysis. Bovine blood was obtained from a healthy donor of CARE-FEPEX ("Cellule d’Appui à la Recherche et à l’Enseignement–Ferme Pédagogique et Experimentale", Dr L. Martinelle) and PBMCs isolated on Ficoll-Paque density gradient using SepMate isolation tubes (STEMCELL technologies).

Myster F., Gong M.J., Javaux J., Suárez N.M., Wilkie G.S., Connelley T., Vanderplasschen A., Davison A.J, & Dewals B.G. (2020). Alcelaphine herpesvirus 1 genes A7 and A8 regulate viral spread and are essential for malignant catarrhal fever. PLoS Pathogens, 16(3), e1008405.

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Isolation and Purification of CD34+ HSPCs

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