Rh il4

Manufactured by Miltenyi Biotec

Sourced in Germany

Rh-IL4 is a recombinant human interleukin-4 (IL-4) protein produced in E. coli. It is used as a cell culture supplement to promote the growth and differentiation of various cell types, such as B cells and T cells.

Automatically generated - may contain errors

Lab products found in correlation

7 protocols using rh il4

Isolation and Activation of Immune Cells

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

PBMCs were isolated from buffy coats from healthy donors (blood transfusion center at University Medical Center (UMC) Mainz, Germany) by Ficoll®-Hypaque (GE Healthcare) density gradient centrifugation. Monocytes were isolated using anti-CD14 microbeads (Miltenyi Biotec) and CD8 and CD4 cells were isolated using anti-CD8 and anti-CD4 microbeads, respectively (Miltenyi Biotec) according to the manufacturer’s protocol. Immature DCs (iDCs) were differentiated from CD14⁺ cells in RPMI 1640 GlutaMAX™, 50 IU/mL penicillin, 50 μg/mL streptomycin, and 10% (v/v) human AB serum (one lambda) (denoted huRPMI), supplemented with 1000 IU/mL recombinant human (rh) GM-CSF and 1000 IU/mL rh IL-4 (Miltenyi Biotec) twice in 5 days. For T-cell activation, 2 × 10⁶ / ml of isolated T-cells were cultivated in 24 well tissue culture plates precoated with 2 μg/ml anti-CD3 antibody (clone OKT3; Bioxcell) in huRPMI supplemented with 100 IU/mL IL-2 for 2 to 3 days. Cells were then harvested, washed and resuspended in huRPMI with 50 IU/mL IL-2 and either rested or immediately submitted to further processing steps.

Alishah K., Birtel M., Masoumi E., Jafarzadeh L., Mirzaee H.R., Hadjati J., Voss R.H., Diken M, & Asad S. (2021). CRISPR/Cas9-mediated TGFβRII disruption enhances anti-tumor efficacy of human chimeric antigen receptor T cells in vitro. Journal of Translational Medicine, 19, 482.

+ Open protocol

+ Expand

Generation of M0-MDM and MDDC from Monocytes

Cited 1 time

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

Unpolarized human monocyte-derived macrophages (M0-MDM) were generated by culturing freshly isolated monocytes in 24-well plates [1×10⁶ cells/well] with CCM [1 mL] containing recombinant human M-CSF (rhM-CSF, Miltenyi Biotec) [50 ng/mL] for three days, replacing half the media [0.5 mL] from each well with fresh CCM [0.5 mL] and rhM-CSF [50 ng/mL], and incubating for another 3 days ^{69 (link)}. Immature human monocytes-derived dendritic cells (MDDC) were generated in the same manner as M0-MDM but incubated with CCM containing recombinant human IL-4 (rhIL-4, Miltenyi Biotec) [50 ng/mL] and recombinant human GM-CSF (Miltenyi Biotec) [160 ng/mL] ^{70 (link)}. On Day 7, media was aspirated, cells were washed once with warm, sterile PBS [37°C], and incubated [37°C / 5%CO₂] with CCM [1 mL] containing the indicated immunostimulant for 24 h.

Alshammari A.M., Smith D.D., Parriott J., Stewart J.P., Curran S.M., McCulloh R.J., Barry P.A., Iyer S.S., Palermo N., Phillips J.A., Dong Y., Ronning D.R., Vennerstrom J.L., Sanderson S.D, & Vetro J.A. (2020). Targeted Amino Acid Substitution Overcomes Scale-Up Challenges with the Human C5a-Derived Decapeptide Immunostimulant EP67. ACS infectious diseases, 6(5), 1169-1181.

+ Open protocol

+ Expand

In vitro Expansion of Human ILC2s

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

Sorted ILC2s (100 cells/well) were expanded with human feeder PBMCs (100,000/well; 37 °C, 5% CO₂) for three to five weeks. Culture medium contained RPMI 1640 Glutamax medium (Life Technologies, Darmstadt, Germany), 1% Pen/Strep (Life Technologies, Darmstadt, Germany), 10% h.i. human AB serum (Sigma-Aldrich, Taufkirchen, Germany), and 25 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, Lonza, Wakersville, USA). The medium was supplemented with 100 U/ml rh-IL-2 (Life Technologies, Darmstadt, Germany), 25 ng/ml rh-IL-4 (Miltenyi Biotech, Bergisch Gladbach, Germany), 5 µg/ml phytohemagglutinin-M (PHA-M; Sigma-Aldrich, Taufkirchen, Germany).

Carstensen S., Gress C., Erpenbeck V.J., Kazani S.D., Hohlfeld J.M., Sandham D.A, & Müller M. (2021). Prostaglandin D2 metabolites activate asthmatic patient-derived type 2 innate lymphoid cells and eosinophils via the DP2 receptor. Respiratory Research, 22, 262.

+ Open protocol

+ Expand

Monocyte-Derived Dendritic Cell Generation

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

Human DCs were derived from monocytes isolated from human peripheral blood supplied by the French Blood Bank (EFS, Rungis, France). Healthy donors gave their written consent for the use of blood donation for research purposes. Peripheral blood mononuclear cells (PBMCs) were sorted from buffy coats by density centrifugation on a Ficoll gradient. Monocytes were then isolated through positive magnetic selection using MidiMacs separation columns and anti-CD14+ antibodies coated on magnetic beads (Miltenyi Biotec, Bergisch Gladbach, Germany). Finally, CD14 cells were differentiated in immature MoDCs for 4 days in RPMI 1640 supplemented with GlutaMAX (Gibco, Invitrogen, Saint Aubin, France), 10 % heat-inactivated fetal calf serum (FCS, Gibco, Invitrogen, Saint Aubin, France), 550 U·mL⁻¹ granulocyte-macrophage colony-stimulating factor (rh-GM-CSF, Miltenyi Biotec, Bergisch Gladbach, Germany), 550 U·mL⁻¹ interleukin-4 (rh-IL4, Miltenyi Biotec, Bergisch Gladbach, Germany), 1 mM sodium pyruvate (Gibco, Invitrogen, Saint Aubin, France), 100 µg·mL⁻¹ streptomycin and 100 U·mL⁻¹ penicillin (Gibco, Invitrogen, Saint Aubin, France).

Feray A., Szely N., Guillet E., Hullo M., Legrand F.X., Brun E., Pallardy M, & Biola-Vidamment A. (2020). How to Address the Adjuvant Effects of Nanoparticles on the Immune System. Nanomaterials, 10(3), 425.

+ Open protocol

+ Expand

Isolation and Differentiation of Human Monocyte-Derived Dendritic Cells

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

Human monocyte-derived dendritic cells (moDCs) were derived from monocytes isolated from human peripheral blood supplied by the French Blood Bank (Établissement Français du Sang, Rungis, France). Healthy donors gave their written consent for the use of blood donation for research purposes. Peripheral blood mononuclear cells (PBMCs) were sorted from buffy coats by density centrifugation on a Ficoll gradient. Monocytes were then isolated through positive magnetic selection using MidiMacs separation columns and anti-CD14 antibodies coated on magnetic beads (Miltenyi Biotec, Bergisch Gladbach, Germany). Finally, CD14⁺ cells were differentiated in immature moDCs for 4 days in RPMI 1640 supplemented with GlutaMAX (Gibco, Invitrogen), 10% heat-inactivated fetal calf serum (FCS, Gibco), 550 U.mL⁻¹ interleukin-4 (rh-IL4, Miltenyi Biotec), 550 U.mL⁻¹ GM-CSF (rh-GM-CSF, Miltenyi Biotech), 1 mM sodium pyruvate (Gibco), 100 µg.mL⁻¹ streptomycin and 100 U.mL⁻¹ penicillin (Gibco) at 37 °C and 5% of CO₂. On day 4, the differentiation of moDCs was evaluated by flow cytometry. The moDCs used will show the following phenotype: CD83 < 5%, CD86 < 30%, CD1a > 80% and DC-SIGN > 90%. moDCs are then used at the density of 1 million per mL.

Guillet É., Brun É., Ferard C., Hardonnière K., Nabhan M., Legrand F.X., Pallardy M, & Biola-Vidamment A. (2023). Human dendritic cell maturation induced by amorphous silica nanoparticles is Syk-dependent and triggered by lipid raft aggregation. Particle and Fibre Toxicology, 20, 12.

+ Open protocol

+ Expand

Isolation and Differentiation of Dendritic Cells and NK Cells

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

Monocytes isolated from HD-PBMCs were sorted as CD14⁺ by using FACSAria cell sorter (BD Bioscience, NJ, USA) and cultured in the presence of rhIL-4 (20 ng/mL, Miltenyi Biotec, Germany) and rhGM-CSF (25 ng/mL, Sargramostim, Leukine©) in RPMI 1640 medium supplemented with 10% fetal bovine serum and with penicillin (100 U/ml) and streptomycin (100 μg/ml). (26 (link)) Following a 6 day of incubation at 37°C, non-adherent cells displayed the CD14^-/CD11c⁺/CD83^- phenotype, which is characteristic of immature DCs. Alternatively, BDCA1⁺DCs were directly isolated from HD or CHB pts at different stage of infection, as Lin-(CD19, CD3, CD14)/HLADR⁺/CD11c⁺/BDCA1⁺.
NK cells were isolated from both HD and CHB pts by negative magnetic separation (NK Cell Isolation Kit/ Miltenyi Biotec, Germany) or sorted as CD3^-/CD56⁺ by FACSAria cell sorter. Cell populations sorted displayed a higher than 95% purity.

De Pasquale C., Campana S., Barberi C., Migliore G.S., Oliveri D., Lanza M., Musolino C., Raimondo G., Ferrone S., Pollicino T, & Ferlazzo G. (2021). Human hepatitis B virus negatively impacts the protective immune cross-talk between natural killer and dendritic cells. Hepatology (Baltimore, Md.), 74(2), 550-565.

+ Open protocol

+ Expand

Generation of Human Monocyte-Derived Dendritic Cells

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

Human monocyte-derived dendritic cells (moDCs) were derived from monocytes isolated from human peripheral blood supplied by the French Blood Bank (EFS, Rungis, France). Healthy donors gave their written consent for the use of blood donation for research purposes. Peripheral blood mononuclear cells (PBMCs) were sorted from buffy coats by density centrifugation on a Ficoll gradient. Monocytes were then isolated through positive magnetic selection using MidiMacs separation columns and anti-CD14+ antibodies coated on magnetic beads (Miltenyi Biotec, Bergisch Gladbach, Germany) . Finally, CD14+ cells were differentiated in immature moDCs for 4 days in RPMI 1640 supplemented with GlutaMAX (Gibco, Invitrogen, Saint Aubin, France), 10 % heat-inactivated fetal bovine serum (FBS, Gibco, Invitrogen, Saint Aubin, France), 550 U.mL -1 granulocyte-macrophage colony-stimulating factor (rh-GM-CSF, Miltenyi Biotec, Bergisch Gladbach, Germany), 550 U.mL -1 interleukin-4 (rh-IL4, Miltenyi Biotec, Bergisch Gladbach, Germany), 1 mM sodium pyruvate (Gibco, Invitrogen, Saint Aubin, France), 100 µg.mL -1 streptomycin and 100 U•mL -1 penicillin (Gibco, Invitrogen, Saint Aubin, France).

Feray A., Guillet É., Szely N., Hullo M., Legrand F.X., Brun E., Rabilloud T., Pallardy M, & Biola-Vidamment A. (2021). Synthetic Amorphous Silica Nanoparticles Promote Human Dendritic Cell Maturation and CD4+ T-Lymphocyte Activation. Toxicological sciences : an official journal of the Society of Toxicology, 185(1).

+ 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!