Neural tissue dissociation kit

Manufactured by Miltenyi Biotec

Sourced in Germany, United States

The Neural Tissue Dissociation Kit is a laboratory equipment designed to efficiently dissociate neural tissue samples into single-cell suspensions. The kit includes reagents and tools necessary for the mechanical and enzymatic dissociation process, enabling the preparation of viable cells from complex neural tissues for further analysis or experimentation.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Tissue dissociation kit (30 citations) Neural Dissociation Kit (26 citations) Neural Tissue Dissociation Kits (P) (3 citations)

182 protocols using neural tissue dissociation kit

Flow Cytometry Analysis of Cellular Markers

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

For all uptake assays cells were analyzed without fixation. For membrane-associated GFP-LC3 analysis, cells were processed as described below. For brain infiltrating monocytes, cells were isolated as described using the Neural Tissue Dissociation Kit (Miltenyi). Primary cells were fixed, permeabilized, and staining using the Cyto Fix/Perm Staining Kit (BD Bioscience) and the indicated, conjugated primary antibodies. For all experiments, cells were analyzed using a Sony SP6800 Spectral Analyzer (Sony). All analyses were performed using FlowJo v10.4 (Tree Star). Fluorescent compensation was performed using BD compensation beads (BD Bioscience).

Heckmann B.L., Teubner B.J., Tummers B., Boada-Romero E., Harris L., Yang M., Guy C.S., Zakharenko S.S, & Green D.R. (2019). LC3-associated endocytosis facilitates β-amyloid clearance and mitigates neurodegeneration in murine Alzheimer’s Disease. Cell, 178(3), 536-551.e14.

+ Open protocol

+ Expand

Murine Glioblastoma Stem Cell Isolation

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

After observing brain tumor formation in an AAV-injected PIC mouse by MRI, the mouse was euthanized and brain tissue was harvested. A portion of the right hemisphere of the brain containing tumor tissue was isolated and dissociated to a single cell suspension using the Neural Tissue Dissociation Kit (Miltenyi) and a gentleMACS Dissociator instrument (Miltenyi). Cells were cultured in 5% CO₂ and 5% O₂ on ultra-low adherence plates to select for neurosphere-forming cells. The resulting murine GSC line was named DF-AA27. Notably, prior attempts to generate murine GSC lines from this GEM model under ambient oxygen conditions were not successful.

Shi D.D., Savani M.R., Levitt M.M., Wang A.C., Endress J.E., Bird C.E., Buehler J., Stopka S.A., Regan M.S., Lin Y.F., Puliyappadamba V.T., Gao W., Khanal J., Evans L., Lee J.H., Guo L., Xiao Y., Xu M., Huang B., Jennings R.B., Bonal D.M., Martin-Sandoval M.S., Dang T., Gattie L.C., Cameron A.B., Lee S., Asara J.M., Kornblum H.I., Mak T.W., Looper R.E., Nguyen Q.D., Signoretti S., Gradl S., Sutter A., Jeffers M., Janzer A., Lehrman M.A., Zacharias L.G., Mathews T.P., Losman J.A., Richardson T.E., Cahill D.P., DeBerardinis R.J., Ligon K.L., Xu L., Ly P., Agar N.Y., Abdullah K.G., Harris I.S., Kaelin WG J.r, & McBrayer S.K. (2022). De novo pyrimidine synthesis is a targetable vulnerability in IDH mutant glioma. Cancer cell, 40(9), 939-956.e16.

+ Open protocol

+ Expand

Isolation and Culture of Primary Microglia

Cited 1 time

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

Cultures were prepared as previously described (Gao et al., 2017 (link)). Briefly, cortices from postnatal day 3–5 C57BL/6 pups were harvested in cold HBSS and enzymatically dissociated into single cell suspensions with a papain-based Neural Tissue Dissociation Kit (Miltenyi) followed by passage through a 70 μm cell strainer. Microglial cells were isolated by positive selection with anti-CD11b magnetic MicroBeads in combination with LS columns (Miltenyi), according to manufacturer’s protocols. Microglia were seeded in poly-L-ornithine-coated 24-well plates (50,000 cells/well) in DMEM/F12 medium supplemented with 20% fetal bovine serum (FBS), 1% MEM Non-Essential Amino Acids, 1% penicillin/streptomycin, 1% GlutaMAX, and 1% Sodium-Pyruvate (all from Gibco-Life Technologies^™). Culture plates were maintained at 37°C in 5% CO₂ atmosphere. After 3 days, the medium was changed and the FBS concentration was reduced to 10%.

Al-Ali H., Gao H., Dalby-Hansen C., Peters V.A., Shi Y, & Brambilla R. (2017). High content analysis of phagocytic activity and cell morphology with PuntoMorph. Journal of neuroscience methods, 291, 43-50.

+ Open protocol

+ Expand

Dissociation and Sorting of Breast Tumors and GBM Xenografts

Cited 1 time

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

4 T1 breast tumors were dissociated into single cell suspensions as previously described [30 (link)]. For GBM xenografts, fresh tissue was finely minced and dissociated into a single cell suspension with the Neural Tissue Dissociation kit (Miltenyi Biotec; Bergisch Gladbach, Germany) according to the manufacturer’s protocols. The mixed cell suspensions were subsequently sorted into isolated tumor and stromal cell populations with the FACS Aria II (Becton Dickinson; Erembodegem, Belgium).

Yang N., Huang B., Tsinkalovsky O., Brekkå N., Zhu H., Leiss L., Enger P.Ø., Li X, & Wang J. (2014). A novel GFP nude rat model to investigate tumor-stroma interactions. Cancer Cell International, 14, 541.

+ Open protocol

+ Expand

Mouse Brain Cell Fractionation

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

Whole brains from adult C57Bl/6 mice were homogenized using an automated homogenizer from Miltenyi Biotec. Solutions used to ensure viability of cells were part of the Neural Tissue Dissociation Kit supplied from Miltenyi Biotec. Following dissociation into a single cell suspension, cells were passed through LS columns (Miltenyi Biotec) containing beads coated with CD11b antibodies (to isolate microglia) or GLAST antibodies (to isolate astrocytes) localized to the columns. The remaining cells not bound to the columns were kept as the neuron enriched fraction. LS columns were washed to remove the CD11b-bound cells or the GLAST-bound cells. The neuronal fraction was stained with CD90-PE antibodies while the microglia fraction was stained with CD11b-PE antibodies. Following staining, cells were analyzed on a BD FACS Canto II flow cytometer (Franklin Lakes, NJ). Data were analyzed using FlowJo Software (Tree Star Incorporated, Ashland, OR).

McMillin M., Frampton G., Tobin R., Dusio G., Smith J., Shin H., Newell-Rogers M.K., Grant S, & DeMorrow S. (2015). TGR5 signaling reduces neuroinflammation during hepatic encephalopathy. Journal of neurochemistry, 135(3), 565-576.

+ Open protocol

+ Expand

Microglial Cell Isolation from Brain

Cited 1 time

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

Brains were collected for cell dissociation using a Neural Tissue Dissociation Kit (Miltenyi Biotec., Bergisch Gladbach, Germany), and microglial cells were then enriched using a magnetic-bead-coupled antibody (anti-CD11b, Miltenyi Biotec, dilution 1:10) extraction technique (MACS™) (Miltenyi Biotec.) according to the manufacturer’s protocol using all recommended reagents and equipment [44 (link)].

Villapol S., Faivre V., Joshi P., Moretti R., Besson V.C, & Charriaut-Marlangue C. (2019). Early Sex Differences in the Immune-Inflammatory Responses to Neonatal Ischemic Stroke. International Journal of Molecular Sciences, 20(15), 3809.

+ Open protocol

+ Expand

Acute Microglia Isolation from Mouse Brain

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

Microglia were acutely isolated from the brain of juvenile or adult mouse without culturing as we reported previously (Jin et al., 2015 (link)). Briefly, brains were dissociated enzymatically with Neural tissue dissociation kit (Miltenyi Biotec, San Diego, CA). Microglia were subsequently purified by the magnetic-activated cell sorting (MACS) using anti-CD11b magnetic beads (Miltenyi Biotec). The whole procedure took about 90 min. Acutely isolated microglia were over 94% pure based on flow cytometry assessments (Jin et al., 2015 (link)).

Horiuchi M., Smith L., Maezawa I, & Jin L.W. (2016). CX3CR1 ablation ameliorates motor and respiratory dysfunctions and improves survival of a Rett syndrome mouse model. Brain, behavior, and immunity, 60, 106-116.

+ Open protocol

+ Expand

Isolation of adult mouse microglia

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

To isolate microglia, adult mice were anesthetized by administrating isoflurane and perfused with ice-cold PBS. The brain was isolated, chopped into fragments, and enzymatically dissociated using a neural tissue dissociation kit (Miltenyi Biotec Inc.). To remove myelin debris, cells were spun down in 30% percoll gradient for 20 min at 600g (accel 5 and decel 1). The myelin layer was carefully discarded, and pellets were filtered through 70 μm strainer and washed two times with 1× HBSS. Subsequently, cells were stained in FACs buffer with targeted antibodies.

Lee J., Villarreal O.D., Wang Y.C., Ragoussis J., Rivest S., Gosselin D, & Richard S. (2022). PRMT1 is required for the generation of MHC-associated microglia and remyelination in the central nervous system. Life Science Alliance, 5(10), e202201467.

+ Open protocol

+ Expand

Isolation and Purification of Brain Cell Types

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

Mice were transcardially perfused with PBS and the brain was removed, digested using a Neural tissue dissociation kit (Miltenyi Biotec, Auburn, CA), and mechanically dissociated with a plastic pipette. After centrifugation at 1000 × g (7 min), the cell pellet was resuspended and incubated with myelin removal beads (Miltenyi Biotec) for 40 min on ice. After washing in PBS, Dynabeads (Thermo Fisher Scientific) conjugated to anti-CD31 (BD Pharmingen, #550274) or anti-CD11b (Biolegend, #127618) were added and a magnetic separator was used to recover the bead-bound cells. Unbound cells were incubated with anti-ACSA + beads (Miltenyi Biotec, #130-097-678) and separated using LS columns (Miltenyi Biotec). Neurons were isolated using a neuron isolation kit (Miltenyi Biotec). Isolated cells were frozen at −80 °C until processing for western blot. We previously reported on the purity of isolated cells by our immunopanning protocol [30 ].

Wu L., Chung J.Y., Cao T., Jin G., Edmiston WJ I.I.I., Hickman S., Levy E.S., Whalen J.A., Abrams E.S., Degterev A., Lo E.H., Tozzi L., Kaplan D.L., El Khoury J, & Whalen M.J. (2021). Genetic inhibition of RIPK3 ameliorates functional outcome in controlled cortical impact independent of necroptosis. Cell Death & Disease, 12(11), 1064.

+ Open protocol

+ Expand

Isolation and characterization of immune cells from the brain

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

Immune cells were isolated from whole-brain homogenates as recently described (15 (link)). Briefly, brain tissue was processed using the neural tissue dissociation kit (Miltenyi Biotec) in accordance with the manufacturer’s specifications. Afterward, cells were separated using a discontinuous Percoll density gradient (30–37–70% Percoll layers), which results in the enrichment of all immune cell types and removes a lot of myelin, which is auto-fluorescent. Following density gradient centrifugation, immune cells were washed and stained for 30 min at 4 °C using the following antibodies: CD11b APC-Cy7 (BD Biosciences), CD45.2 Pacific blue, CD86 PE-Cy5, and CD206 PE-Cy7 (BioLegend). Respective isotype control antibodies or unstained samples were used to determine positive populations. Myelin debris and dead cells were excluded by FSC/SSC gating, and singlet populations were analyzed.
For flow cytometric analyses of blood samples, EDTA-blood was stained at 4 °C for 15 min with the following antibodies: B220 Pacific Blue, CD3 AF700, CSF-1R BV605 (BioLegend), and CD11b APC-Cy7 (BD Biosciences). For more details see Waltl et al. (15 (link)).
Flow cytometry was performed using an LSRII flow cytometer (BD Biosciences), and data were analyzed using FlowJo software (Tree Star).

Käufer C., Chhatbar C., Bröer S., Waltl I., Ghita L., Gerhauser I., Kalinke U, & Löscher W. (2018). Chemokine receptors CCR2 and CX3CR1 regulate viral encephalitis-induced hippocampal damage but not seizures. Proceedings of the National Academy of Sciences of the United States of America, 115(38), E8929-E8938.

+ 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!