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Il 34

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IL-34 is a lab equipment product from Thermo Fisher Scientific. It is used for the detection and quantification of interleukin-34 (IL-34), a cytokine involved in various biological processes. The core function of IL-34 is to provide a reliable and accurate method for researchers to measure IL-34 levels in their samples.

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Lab products found in correlation

M csf, by Thermo Fisher Scientific (22 mentions) Glutamax, by Thermo Fisher Scientific (18 mentions) Cx3cl1, by Thermo Fisher Scientific (10 mentions) Tgf β1, by Thermo Fisher Scientific (9 mentions) Gm csf, by Thermo Fisher Scientific (8 mentions) Dmem f12, by Thermo Fisher Scientific (8 mentions) Cd200, by Novoprotein (7 mentions) Stemdiff hematopoietic kit, by STEMCELL (6 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions) X vivo 15, by Lonza (4 mentions) Stem cell factor (scf), by Thermo Fisher Scientific (4 mentions) Interleukin 3 (il 3), by Thermo Fisher Scientific (4 mentions) Vascular endothelial growth factor (vegf), by Thermo Fisher Scientific (4 mentions) Stemdiff hematopoesis kit, by STEMCELL (3 mentions) Tgf β2, by Thermo Fisher Scientific (3 mentions) Poly d lysine, by Merck Group (3 mentions) β mercaptoethanol, by Thermo Fisher Scientific (3 mentions) Insulin, by Merck Group (3 mentions) 2 mercaptoethanol, by Thermo Fisher Scientific (3 mentions) N2 supplement, by Thermo Fisher Scientific (3 mentions)

Close spelling variants of this product

Human IL-34 (2 citations)

33 protocols using il 34

1

Modulating Monocyte Differentiation

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To further analyze the role of CD115 signaling during monocyte differentiation, 100 ng/mL of their ligands, CSF1 and IL34 (both from Peprotech), was added independently or in combination to the culture of mDC and tolDC on day 0 and day 4. Phenotype and functional characterization of CSF1- and/or IL34-treated cells were analyzed on day 6.
Mansilla M.J., González-Larreategui I., Figa-Martín N., Barallat J., Fondelli F., Sellés-Rius A., Quirant-Sánchez B., Teniente-Serra A, & Martínez-Cáceres E. (2021). Transfection of Vitamin D3-Induced Tolerogenic Dendritic Cells for the Silencing of Potential Tolerogenic Genes. Identification of CSF1R-CSF1 Signaling as a Glycolytic Regulator. International Journal of Molecular Sciences, 22(14), 7363.
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2

Differentiation of Microglia Precursors

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The 50 K macrophage precursors were plated per well in a glass‐bottom 96‐glass bottom well plate (IBL Baustoff, 220.230.042). Cells were cultured with microglia differentiation medium (Advanced DMEM/F12 [Thermo Fisher, 12634010], 1× N2 [Thermo Fisher, 17502001], 1× Pen/Strep [Invitrogen, 15140122], 1× GlutaMax [Thermo Fisher, 35050061], 50 μM 2‐mercaptoethanol [Thermo Fisher, 31350‐010], 100 ng/ml IL‐34 [Peprotech, 200‐34], 10 ng/ml GM‐CSF [Peprotech, 300‐03]). Cells were kept in culture for 10 days, with a medium change every 3–4 days, and then fixed with 4% formaldehyde (Millipore, 1.00496.5000) for immunostaining or used for a phagocytosis or MTT assay.
Sabate‐Soler S., Nickels S.L., Saraiva C., Berger E., Dubonyte U., Barmpa K., Lan Y.J., Kouno T., Jarazo J., Robertson G., Sharif J., Koseki H., Thome C., Shin J.W., Cowley S.A, & Schwamborn J.C. (2022). Microglia integration into human midbrain organoids leads to increased neuronal maturation and functionality. Glia, 70(7), 1267-1288.
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3

Differentiation of iHPCs and iPS-microglia

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iHPCs and iPS-microglia were differentiated according to the protocol published by McQuade et al. (McQuade et al., 2018 (link)). To begin iHPC differentiation, iPSCs were passaged in mTeSR-E8 to achieve a density of 80 colonies of 100 cells each per 35mm well. On day 0, cells were transferred to Medium A from the STEMdiff™ Hematopoietic Kit (Stem Cell Technologies). On day 3, flattened endothelial cell colonies were exposed to Medium B and cells remained in medium B for 7 additional days while iHPCs began to lift off the colonies. On day 10, non-adherent CD43+ iHPCs were collected by removing medium and cells with a serological pipette. At this point, d10-d11 iHPCs can be frozen in Bambanker (Wako). Cells used for early-postnatal iHPC transplantation were thawed in iPS-Microglia medium (DMEM/F12, 2X insulin-transferrin-selenite, 2X B27, 0.5X N2, 1X glutamax, 1X non-essential amino acids, 400 μM monothioglycerol, and 5 μg/mL human insulin freshly supplemented with 100ng/mL IL-34, 50ng/mL TGFβ1, and 25 ng/mL M-CSF (Peprotech)) and allowed to recover for 24 hours, then resuspended at 62,500 cells/uL in 1X DPBS (low Ca2+, low Mg2+). Cells utilized for in vitro experiments continued microglial differentiation for 28 days. During the last 3 days in culture, 100ng/mL CD200 (Novoprotein) and 100 ng/mL CX3CL1 (Peprotech) were added to mature microglia in a brain-like environment.
Hasselmann J., Coburn M.A., England W., Velez D.X., Shabestari S.K., Tu C.H., McQuade A., Kolahdouzan M., Echeverria K., Claes C., Nakayama T., Azevedo R., Coufal N.G., Han C.Z., Cummings B.J., Davtyan H., Glass C.K., Healy L.M., Gandhi S.P., Spitale R.C, & Blurton-Jones M. (2019). Development of a chimeric model to study and manipulate human microglia in vivo. Neuron, 103(6), 1016-1033.e10.
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4

Generating iPSC-derived Microglia

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iPSC-microglia were generated as described in McQuade et al., 2018 (link) and McQuade and Blurton-Jones, 2021 (link). Briefly, iPSCs were directed down a hematopoietic lineage using the STEMdiff Hematopoietic kit (STEMCELL Technologies). After 10–12 days in culture, CD43+ hematopoietic progenitor cells were transferred into a microglia differentiation medium containing DMEM/F12, 2× insulin-transferrin-selenite, 2× B27, 0.5× N2, 1× GlutaMAX, 1× nonessential amino acids, 400  μM monothioglycerol, and 5  μg/mL human insulin. Media was added to cultures every other day and supplemented with 100  ng/mL IL-34, 50  ng/mL TGF-β1, and 25 ng/mL M-CSF (PeproTech) for 28 days. In the final 3 days of differentiation, 100  ng/mL CD200 (Novoprotein) and 100  ng/mL CX3CL1 (PeproTech) were added to culture.
Jairaman A., McQuade A., Granzotto A., Kang Y.J., Chadarevian J.P., Gandhi S., Parker I., Smith I., Cho H., Sensi S.L., Othy S., Blurton-Jones M, & Cahalan M.D. (2022). TREM2 regulates purinergic receptor-mediated calcium signaling and motility in human iPSC-derived microglia. eLife, 11, e73021.
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5

Microglia Differentiation from iPSCs

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iPSCs were differentiated into microglia following an established protocol75 (link) with minor changes. In brief, embryoid bodies (EBs) were formed using AggreWell™800 (STEMCELL Technologies), and cultured in mTeSR1 (STEMCELL Technologies) with bone morphogenetic protein 4 (BMP4, 50 ng/mL; ImmunoTools), vascular endothelial growth factor (VEGF, 50 ng/mL; ImmunoTools), and stem cell factor (SCF, 20 ng/mL; ImmunoTools) for four days with 75% medium change daily. On day 4, EBs were collected and transferred into 6-well cell culture plates (12–16 EBs/well) in X-VIVO 15 (Lonza) supplemented with IL-3, (25 ng/mL; ImmunoTools), macrophage colony-stimulating factor (M-CSF, 100 ng/mL; ImmunoTools), 2 mM GlutaMAX (Thermo Fisher), 1% P/S (Merck Millipore), and 0.055 mM β-mercaptoethanol (Sigma-Aldrich), with medium change weekly. After 3–4 weeks, floating cells were collected and seeded at a concentration of 100,000 cells/cm2 on Matrigel-coated plates in Advanced DMEM/F-12 (Life Technologies) supplemented with N2 (Thermo Fisher), GlutaMAX, P/S, β-mercaptoethanol, M-CSF (100 ng/mL), IL-34 (100 ng/mL; PeproTech), and GM-CSF (10 ng/mL, ImmunoTools) with medium change twice a week.
Panagiotakopoulou V., Ivanyuk D., De Cicco S., Haq W., Arsić A., Yu C., Messelodi D., Oldrati M., Schöndorf D.C., Perez M.J., Cassatella R.P., Jakobi M., Schneiderhan-Marra N., Gasser T., Nikić-Spiegel I, & Deleidi M. (2020). Interferon-γ signaling synergizes with LRRK2 in neurons and microglia derived from human induced pluripotent stem cells. Nature Communications, 11, 5163.
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6

Monocyte-derived Macrophage-like Cells Generation

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Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood of HIV seronegative donors, and monocytes prepared as described above. MMG were generated from isolated monocytes using methods modified from those previously described [34 (link), 37 (link)]. To induce the differentiation of MMG, monocytes were cultured in RPMI-1640 supplemented with 1X GlutaMAX (Life Technologies), 1% penicillin/streptomycin and a mixture of the following human recombinant cytokines: M-CSF (10 ng/mL; Peprotech), GM-CSF (10 ng/mL; Peprotech), NGF-β (10 ng/mL; Peprotech), CCL2 (100 ng/mL; Peprotech), and IL-34 (100 ng/mL; Peprotech) under standard humidified culture conditions (37 °C, 5% CO2). The monocytes were incubated with 1% serum for the first 3 days, and thereafter, every third day, cells were supplemented with fresh serum-free media containing M-CSF, GM-CSF, NGF-β, IL-34 and CCL2. Cells were cultured for 14 days. Characterization experiments were performed on day 14.
Rai M.A., Hammonds J., Pujato M., Mayhew C., Roskin K, & Spearman P. (2020). Comparative analysis of human microglial models for studies of HIV replication and pathogenesis. Retrovirology, 17, 35.
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7

Differentiation of Microglia from hiPSCs

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Details of microglial differentiation from hiPSCs are summarized in Sonn et al., 2022 (link). Briefly, GeneJuice (Sigma–Aldrich, 70,967) and Opti-MEM medium (Gibco™, 31,985,070) were used to establish a doxycycline-inducible PU.1-overexpression clone of hiPSCs. Then, the generation of microglial cells from iPSCs was divided into two stages. First, hematopoietic progenitor cells were generated from hiPSCs (hiHPCs; ∼day 18) in modified StemPro™-34 SFM (1×) (Gibco™, 10,639,011) medium supplied with multiple cytokines and chemicals (BMP4 (PEPROTECH), CHIR99021 (Focus Biomolecules), VEGF (Thermo Fisher Scientific), FGF2 (PEPROTECH), SCF (PEPROTECH), IL-3 (PEPROTECH), IL-6 (PEPROTECH), and IWR-1e (Thermo Fisher Scientific). Doxycycline (1 μg/mL) was added to the medium during days 6 ∼ 18.
Second, microglia-like cells were differentiated from hiHPCs (day 19∼) in modified DMEM/F12 (1:1, Gibco™) supplied with a cocktail of five cytokines (100 ng/mL IL-34, 50 ng/mL TGFβ1, 25 ng/mL M-CSF, 100 ng/mL CD200, and 100 ng/mL CX3CL1, all from PEPROTECH). Induced microglia were used for analysis on day 21.
Kase Y., Sonn I., Goto M., Murakami R., Sato T, & Okano H. (2023). The original strain of SARS-CoV-2, the Delta variant, and the Omicron variant infect microglia efficiently, in contrast to their inability to infect neurons: Analysis using 2D and 3D cultures. Experimental Neurology, 363, 114379.
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8

Differentiation of Macrophage Precursors

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PMPs were differentiated in the medium composed of DMEM/ F12
supplemented with N2, 2 mM Glutamax, 100 U/mL penicillin and 100 mg/mL
streptomycin, 100 ng/mL M-CSF (Peprotech), 100 ng/mL IL-34 (Peprotech), and
10 ng/mL GM-CSF (Peprotech) for two weeks (Haenseler et al., 2017 (link)). The medium was changed once a week.
After two weeks, cells were collected for Western blotting.
Jin M., Xu R., Wang L., Alam M.M., Ma Z., Zhu S., Martini A.C., Jadali A., Bernabucci M., Xie P., Kwan K.Y., Pang Z.P., Head E., Liu Y., Hart R.P, & Jiang P. (2022). Type I Interferon Signaling Drives Microglial Dysfunction and Senescence in Human iPSC Models of Down Syndrome and Alzheimer’s Disease. Cell stem cell, 29(7), 1135-1153.e8.
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9

Generating iMGL from Human iPSCs

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A detailed step-by-step protocol for generation of iMGL from iPSCs can be found in the supplementary materials (Supplementary File S1). In brief, human iPSCs were seeded as small clumps and differentiated into HPCs using the STEMDiff hematopoietic kit (STEMCELL Technologies Vancouver, BC, Canada) with the extension of an additional harvesting step on day 14 of culture using RPMI1640 + 10% FCS (Gibco, Waltham, MA, USA) + 10 ng/mL GM-CSF (Peprotech, Cranbury, NJ, USA) + Penicillin/Streptomycin which are not provided with the kit. HPCs harvested on day 12 and 14 were pooled and either cryopreserved or directly used for further maturation into iMGL. For maturation, HPCs were seeded in maturation medium (RPMI1640 + 10% FCS (Gibco, Waltham, MA, USA) + Penicillin/Streptomycin + 100 ng/mL IL-34 (Peprotech, Cranbury, NJ, USA) + 10 ng/mL GM-CSF (Peprotech, Cranbury, NJ, USA)) at a density of 90.000–100.000 cells/cm2 onto glass slides or the different coating modalities depicted in the respective experiments. Maturation medium was added every 2–3 days for 2 weeks with a half medium change after 1 week. After 2 weeks, iMGL were used for subsequent experiments.
Lanfer J., Kaindl J., Krumm L., Gonzalez Acera M., Neurath M., Regensburger M., Krach F, & Winner B. (2022). Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells. International Journal of Molecular Sciences, 23(9), 4526.
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10

Directed Differentiation of iPSCs to Microglia

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iPSC-microglia were generated as described (McQuade et al., 2018 (link)). Briefly, iPSCs were directed down a hematopoietic lineage using the STEMdiff Hematopoesis kit (StemCell Technologies). After 10–12 days in culture, CD43+ hematopoteic progenitor cells are transferred into a microglia differentiation medium containing DMEM/F12, 2× insulin-transferrin-selenite, 2× B27, 0.5× N2, 1× glutamax, 1× non-essential amino acids, 400 µM monothioglycerol, and 5 µg/mL human insulin. Media was added to cultures every other day and supplemented with 100 ng/mL IL-34, 50 ng/mL TGF-β1, and 25 ng/mL M-CSF (Peprotech) for 28 days. In the final 3 days of differentiation 100 ng/mL CD200 (Novoprotein) and 100 ng/mL CX3CL1 (Peprotech) were added to culture.
Shabestari S.K., Morabito S., Danhash E.P., McQuade A., Sanchez J.R., Miyoshi E., Chadarevian J.P., Claes C., Coburn M.A., Hasselmann J., Hidalgo J., Tran K.N., Martini A.C., Rothermich W.C., Pascual J., Head E., Hume D.A., Pridans C., Davtyan H., Swarup V, & Blurton-Jones M. (2022). Absence of microglia promotes diverse pathologies and early lethality in Alzheimer’s disease mice. Cell reports, 39(11), 110961.
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