Isolation of murine microglial cells for RNA analysis or flow cytometry

Cardona, Astrid E; Huang, DeRen; Sasse, Margaret E; Ransohoff, Richard M

doi:10.1038/nprot.2006.327

Cited by 236Neuroinflammation and Neurodegeneration MechanismsNeurogenesis and neuroplasticity mechanismsNeurological Disease Mechanisms and Treatments

Isolation of murine microglial cells for RNA analysis or flow cytometry

Cardona, Astrid E, Huang, DeRen, Sasse, Margaret E, Ransohoff, Richard M·2006·Nature Protocols

DOI 10.1038/nprot.2006.327

Open this paper in CoChat — free account

Abstract

There is increasing interest in the isolation of adult microglia to study their functions at a morphological and molecular level during normal and neuroinflammatory conditions. Microglia have important roles in brain homeostasis, and in disease states they exert neuroprotective or neurodegenerative functions. To assay expression profiles or functions of microglia, we have developed a method to isolate microglial cells and infiltrating leukocytes from adult mouse brain. This protocol uses a digestion cocktail containing collagenase and dispase, and it involves separation over discontinuous percoll gradients. Isolated cells can be used for RNA analysis, including RNase protection analysis (RPA), quantitative RT-PCR, high-density microarray, proteomic or flow cytometric characterization of cell surface markers or adoptive transfer. Cell isolation can be completed in less than 4 h.

Integrity signals

Registry

Crossref · 10.1038/nprot.2006.327

Cites

35 of 35 references verified0 citation errors · 0 not checkable

Statistics

Recomputing p-values needs the paper’s full text.

Claims the paper makes

This paper’s full text isn’t openly available, so its claims and self-check weren’t run. The registry and citation checks still apply.

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Previous reports comparing cell isolation protocols or cells postisolation have often focused on aspects such as cell viability and/or yield over whether the isolated cells are truly reflective of their in vivo state.
“Previous reports comparing cell isolation protocols or cells postisolation have often focused on aspects such as cell viability and/or yield4,62 over whether the isolated cells are truly reflective of their in vivo state13,14.”
From Dissection of artifactual and confounding glial signatures by single-cell sequencing of mouse and human brain (2022) · cites this paper as [4]
Many widely used protocols for microglial isolation continue to include enzymatic digestion, likely because some experimental conditions require greater yield.
“Some experimental conditions may dictate the need for greater yield, especially for older or less-sensitive approaches (including many older bulk RNA-sequencing (RNA-seq) studies or protein-based analyses), which is likely why many widely used and widely cited protocols for microglial isolation continue to include enzymatic digestion4,63,64.”
From Dissection of artifactual and confounding glial signatures by single-cell sequencing of mouse and human brain (2022) · cites this paper as [4]
Anti-FCRLS and anti-P2ry12 antibodies stain adult microglia isolated from naïve murine brain but did not stain CD11b-gated myeloid cells from murine spleen, bone marrow and peripheral blood.
“FACS analysis shows that anti-FCRLS and anti-P2ry12 antibodies stain adult microglia isolated by conventional methods22 from naïve murine brain but did not stain CD11b-gated myeloid cells from murine spleen, bone marrow and peripheral blood (Fig. 4a).”
From Identification of a unique TGF-β–dependent molecular and functional signature in microglia (2014) · cites this paper as [22]

Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo
Nimmerjahn, A., Kirchhoff, F., & Helmchen, F. (2005) · Science · doi:10.1126/science.1110647 · Cited by 5,827 · see citing works
ATP mediates rapid microglial response to local brain injury in vivo
Davalos, D., Grutzendler, J., Yang, G., Kim, J. V., Zuo, Y., Jung, S., Littman, D. R., Dustin, M. L., & Gan, W.-B. (2005) · Nature Neuroscience · doi:10.1038/nn1472 · Cited by 3,913 · see citing works
Analysis of Fractalkine Receptor CX<sub>3</sub>CR1 Function by Targeted Deletion and Green Fluorescent Protein Reporter Gene Insertion
Jung, S., Aliberti, J., Graemmel, P., Sunshine, M. J., Kreutzberg, G. W., Sher, A., & Littman, D. R. (2000) · Molecular and Cellular Biology · doi:10.1128/mcb.20.11.4106-4114.2000 · Cited by 2,632 · see citing works
Control of microglial neurotoxicity by the fractalkine receptor
Cardona, A. E., Pioro, E. P., Sasse, M. E., Kostenko, V., Cardona, S. M., Dijkstra, I. M., Huang, D., Kidd, G., Dombrowski, S., Dutta, R., et al. (2006) · Nature Neuroscience · doi:10.1038/nn1715 · Cited by 1,531 · see citing works
Self-citation
Immune function of microglia
Aloisi, F. (2001) · Glia · doi:10.1002/glia.1106 · Cited by 1,281 · see citing works
Cytology & cellular pathology of the nervous system
Penfield, W. G. (1965) · Medical Entomology and Zoology · Cited by 1,035 · see citing works
Microglia as neuroprotective, immunocompetent cells of the CNS
Streit, W. J. (2002) · Glia · doi:10.1002/glia.10154 · Cited by 919 · see citing works
Experimental autoimmune encephalomyelitis repressed by microglial paralysis
Heppner, F. L., Greter, M., Marino, D., Falsig, J., Raivich, G., Hövelmeyer, N., Waisman, A., Rülicke, T., Prinz, M., Priller, J., et al. (2005) · Nature Medicine · doi:10.1038/nm1177 · Cited by 732 · see citing works
Isolation and direct characterization of resident microglial cells from the normal and inflamed central nervous system.
Sedgwick, J. D., Schwender, S., Imrich, H., Dörries, R., Butcher, G. W., & ter Meulen, V. (1991) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.88.16.7438 · Cited by 691 · see citing works
Normal adult ramified microglia separated from other central nervous system macrophages by flow cytometric sorting. Phenotypic differences defined and direct ex vivo antigen presentation to myelin basic protein-reactive CD4+ T cells compared
Ford, A. L., Goodsall, A. L., Hickey, W. F., & Sedgwick, J. D. (1995) · The Journal of Immunology · doi:10.4049/jimmunol.154.9.4309 · Cited by 631 · see citing works
Dynamics of microglial activation: A confocal time-lapse analysis in hippocampal slices
Stence, N., Waite, M., & Dailey, M. E. (2001) · Glia · doi:10.1002/1098-1136(200103)33:3 · Cited by 617 · see citing works
The role of human glioma-infiltrating microglia/macrophages in mediating antitumor immune responses1
Hussain, S. F., Yang, D., Suki, D., Aldape, K., Grimm, E., & Heimberger, A. B. (2006) · Neuro-Oncology · doi:10.1215/15228517-2006-008 · Cited by 592 · see citing works
Macrophages and the Nervous System
Perry, V. H., & Gordon, S. (1991) · International review of cytology · doi:10.1016/s0074-7696(08)61220-6 · Cited by 428 · see citing works
Microglial cell activation and proliferation precedes the onset of CNS autoimmunity
Ponomarev, E. D., Shriver, L. P., Maresz, K., & Dittel, B. N. (2005) · Journal of Neuroscience Research · doi:10.1002/jnr.20488 · Cited by 349 · see citing works
The neuronal chemokine CX3CL1/fractalkine selectively recruits NK cells that modify experimental autoimmune encephalomyelitis within the central nervous system
Huang, D., Shi, F., Jung, S., Pien, G. C., Wang, J., Salazar‐Mather, T. P., He, T. T., Weaver, J. T., Ljunggren, H., Biron, C. A., et al. (2006) · The FASEB Journal · doi:10.1096/fj.05-5465com · Cited by 284 · see citing works
Self-citation
How the Blood Talks to the Brain Parenchyma and the Paraventricular Nucleus of the Hypothalamus During Systemic Inflammatory and Infectious Stimuli
Rivest, S., Lacroix, S., Vallières, L., Nadeau, S., Zhang, J., & Laflamme, N. (2000) · Proceedings of The Society for Experimental Biology and Medicine · doi:10.1046/j.1525-1373.2000.22304.x · Cited by 270 · see citing works
Microglia as liaisons between the immune and central nervous systems: Functional implications for multiple sclerosis
Carson, M. J. (2002) · Glia · doi:10.1002/glia.10145 · Cited by 227 · see citing works
Cytology and Cellular Pathology of the Nervous System.
(1932) · JAMA · doi:10.1001/jama.1932.02740560055030 · Cited by 214 · see citing works
IL-23 produced by CNS-resident cells controls T cell encephalitogenicity during the effector phase of experimental autoimmune encephalomyelitis
Becher, B., Durell, B. G., & Noelle, R. J. (2003) · Journal of Clinical Investigation · doi:10.1172/jci200319079 · Cited by 173 · see citing works
Inverted Immunodominance and Impaired Cytolytic Function of CD8+ T Cells During Viral Persistence in the Central Nervous System
Bergmann, C. C., Altman, J. D., Hinton, D. R., & Stohlman, S. A. (1999) · The Journal of Immunology · doi:10.4049/jimmunol.163.6.3379 · Cited by 172 · see citing works
A Mechanism for Neurodegeneration Induced by Group B Streptococci through Activation of the TLR2/MyD88 Pathway in Microglia
Lehnardt, S., Henneke, P., Lien, E., Kasper, D. L., Volpe, J. J., Bechmann, I., Nitsch, R., Weber, J. R., Golenbock, D. T., & Vartanian, T. K. (2006) · The Journal of Immunology · doi:10.4049/jimmunol.177.1.583 · Cited by 171 · see citing works
IL-23 produced by CNS-resident cells controls T cell encephalitogenicity during the effector phase of experimental autoimmune encephalomyelitis
Becher, B., Durell, B. G., & Noelle, R. J. (2003) · Journal of Clinical Investigation · doi:10.1172/jci19079 · Cited by 165 · see citing works
Resident and Infiltrating Central Nervous System APCs Regulate the Emergence and Resolution of Experimental Autoimmune Encephalomyelitis
Juedes, A. E., & Ruddle, N. H. (2001) · The Journal of Immunology · doi:10.4049/jimmunol.166.8.5168 · Cited by 145 · see citing works
Central Role for MyD88 in the Responses of Microglia to Pathogen-Associated Molecular Patterns
Esen, N., & Kielian, T. (2006) · The Journal of Immunology · doi:10.4049/jimmunol.176.11.6802 · Cited by 107 · see citing works
Long term culture of bovine oligodendroglia isolated with a percoll gradient
Lisak, R. P., Pleasure, D., Silberberg, D. H., et al. (1981) · Brain Research · doi:10.1016/0006-8993(81)90809-x · Cited by 86 · see citing works
Expression of two temporally distinct microglia‐related gene clusters after spinal cord injury
Byrnes, K., Garay, J., Giovanni, S. D., Biase, A. D., Knoblach, S., Hoffman, E., Movsesyan, V., & Faden, A. I. (2006) · Glia · doi:10.1002/glia.20295 · Cited by 73 · see citing works
Pertussis Toxin-Induced Reversible Encephalopathy Dependent on Monocyte Chemoattractant Protein-1 Overexpression in Mice
Huang, D., Tani, M., Wang, J., Han, Y., He, T. T., Weaver, J. T., Charo, I. F., Tuohy, V. K., Rollins, B. J., & Ransohoff, R. M. (2002) · Journal of Neuroscience · doi:10.1523/jneurosci.22-24-10633.2002 · Cited by 60 · see citing works
Self-citation
Towards a transcriptome definition of microglial cells
Moran, L. B., Duke, D. C., Turkheimer, F., Banati, R. B., & Graeber, M. B. (2004) · Neurogenetics · doi:10.1007/s10048-004-0172-5 · Cited by 59 · see citing works
Microarray analysis of activated mixed glial (microglia) and monocyte-derived macrophage gene expression
Albright, A. V., & González‐Scarano, F. (2004) · Journal of Neuroimmunology · doi:10.1016/j.jneuroim.2004.09.007 · Cited by 57 · see citing works
Transgenic expression of CCL2 in the central nervous system prevents experimental autoimmune encephalomyelitis
Elhofy, A., Wang, J., Tani, M., Fife, B. T., Kennedy, K. J., Bennett, J. L., Huang, D., Ransohoff, R. M., & Karpus, W. J. (2004) · Journal of Leukocyte Biology · doi:10.1189/jlb.0804465 · Cited by 42 · see citing works
Self-citation
Anatomical Gradients in Proliferation and Differentiation of Embryonic Rat CNS Accessed by Buoyant Density Fractionation: α3, β3 and γ2 GABA A Receptor Subunit Co‐expression by Post‐mitotic Neocortical Neurons Correlates Directly with Cell Buoyancy
Maric, D., Marić, I., Ma, W., et al. (1997) · European Journal of Neuroscience · doi:10.1111/j.1460-9568.1997.tb01628.x · Cited by 40 · see citing works
Isolation of Hippocampal Synaptosomes on Percoll Gradients: Cholinergic Markers and Ligand Binding Sites
Thorne, B., Wonnacott, S., & Dunkley, P. R. (1991) · Journal of Neurochemistry · doi:10.1111/j.1471-4159.1991.tb08175.x · Cited by 31 · see citing works
Microarrays and expression profiling in microglia research and in inflammatory brain disorders
Gebicke‐Haerter, P. J. (2005) · Journal of Neuroscience Research · doi:10.1002/jnr.20479 · Cited by 31 · see citing works
Amylose bronchique localisée : Une observation
Ayadi, L., Khabir, A., Boudawara, T. S., et al. (2004) · Annales de Pathologie · doi:10.1016/s0242-6498(04)93897-7 · Cited by 5 · see citing works
Neurotoxicité et neuroprotection, les deux facettes de l’activation microgliale au cours de l’infection par le virus de l’immunodéficience humaine (VIH)
Vallat‐Decouvelaere, A., Gray, F., Chrétien, F. C., et al. (2004) · Annales de Pathologie · doi:10.1016/s0242-6498(04)93895-3 · Cited by 5 · see citing works

Flagged first, then by citations

Sorted by citations

Identification of a unique TGF-β–dependent molecular and functional signature in microglia
Butovsky, O., Jedrychowski, M. P., Moore, C. S., Cialic, R., Lanser, A. J., Gabriely, G., Koeglsperger, T., Dake, B., Wu, P. M., Doykan, C. E., et al. (2014) · Nature Neuroscience · doi:10.1038/nn.3599 · Cited by 2,543 · see citing works
Self-citation
Microglial physiology: unique stimuli, specialized responses
Ransohoff, R. M., & Perry, V. H. (2009) · Annual Review of Immunology · doi:10.1146/annurev.immunol.021908.132528 · Cited by 1,858 · see citing works
Self-citation
A Neurodegeneration-Specific Gene-Expression Signature of Acutely Isolated Microglia from an Amyotrophic Lateral Sclerosis Mouse Model
Chiu, I. M., Morimoto, E. T. A., Goodarzi, H., Liao, J. T., O’Keeffe, S., Phatnani, H. P., Muratet, M., Carroll, M. C., Levy, S., Tavazoie, S., et al. (2013) · Cell Reports · doi:10.1016/j.celrep.2013.06.018 · Cited by 651 · see citing works
Microglia dictate the impact of saturated fat consumption on hypothalamic inflammation and neuronal function.
Valdearcos, M., Robblee, M. M., Benjamin, D. I., Nomura, D. K., Xu, A. W., & Koliwad, S. K. (2014) · Cell Reports · doi:10.1016/j.celrep.2014.11.018 · Cited by 629 · see citing works
Selective chemokine receptor usage by central nervous system myeloid cells in CCR2-red fluorescent protein knock-in mice
Saederup, N., Cardona, A. E., Croft, K., Mizutani, M., Cotleur, A. C., Tsou, C.-L., Ransohoff, R. M., & Charo, I. F. (2010) · PLoS ONE · doi:10.1371/journal.pone.0013693 · Cited by 599 · see citing works
Self-citation
Peripheral lipopolysaccharide (LPS) challenge promotes microglial hyperactivity in aged mice that is associated with exaggerated induction of both pro-inflammatory IL-1β and anti-inflammatory IL-10 cytokines
Henry, C. J., Huang, Y., Wynne, A. M., & Godbout, J. P. (2009) · Brain, Behavior, and Immunity · doi:10.1016/j.bbi.2008.09.002 · Cited by 565 · see citing works
Microglial inflammatory signaling orchestrates the hypothalamic immune response to dietary excess and mediates obesity susceptibility.
Valdearcos, M., Douglass, J. D., Robblee, M. M., Dorfman, M. D., Stifler, D. R., Bennett, M. L., Gerritse, I., Fasnacht, R., Barres, B. A., Thaler, J. P., et al. (2017) · Cell Metabolism · doi:10.1016/j.cmet.2017.05.015 · Cited by 477 · see citing works
Dissection of artifactual and confounding glial signatures by single-cell sequencing of mouse and human brain
Marsh, S. E., Walker, A. J., Kamath, T., Dissing-Olesen, L., Hammond, T. R., de Soysa, T. Y., Young, A. M. H., Murphy, S., Abdulraouf, A., Nadaf, N., et al. (2022) · Nature Neuroscience · doi:10.1038/s41593-022-01022-8 · Cited by 432 · see citing works
Infiltrating monocytes promote brain inflammation and exacerbate neuronal damage after status epilepticus
Varvel, N. H., Neher, J. J., Bosch, A., Wang, W., Ransohoff, R. M., Miller, R. J., & Dingledine, R. (2016) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1604263113 · Cited by 357 · see citing works
Self-citation
Untangling the brain’s neuroinflammatory and neurodegenerative transcriptional responses
Srinivasan, K., Friedman, B. A., Larson, J. L., Lauffer, B. E., Goldstein, L. D., Appling, L. L., Borneo, J., Poon, C., Ho, T., Cai, F., et al. (2016) · Nature Communications · doi:10.1038/ncomms11295 · Cited by 336 · see citing works
Transcriptional signature in microglia associated with Aβ plaque phagocytosis
Grubman, A., Choo, X. Y., Chew, G., Ouyang, J. F., Sun, G., Croft, N. P., Rossello, F. J., Simmons, R., Buckberry, S., Landin, D. V., et al. (2021) · Nature Communications · doi:10.1038/s41467-021-23111-1 · Cited by 357 · see citing works
The fractalkine receptor but not CCR2 is present on microglia from embryonic development throughout adulthood
Mizutani, M., Pino, P. A., Saederup, N., Charo, I. F., Ransohoff, R. M., & Cardona, A. E. (2012) · The Journal of Immunology · doi:10.4049/jimmunol.1100421 · Cited by 346 · see citing works
Self-citation
Single-cell mass cytometry reveals distinct populations of brain myeloid cells in mouse neuroinflammation and neurodegeneration models
Ajami, B., Samusik, N., Wieghofer, P., Ho, P. P., Crotti, A., Bjornson, Z., Prinz, M., Fantl, W. J., Nolan, G. P., & Steinman, L. (2018) · Nature Neuroscience · doi:10.1038/s41593-018-0100-x · Cited by 338 · see citing works
Antibiotic-induced perturbations in microbial diversity during post-natal development alters amyloid pathology in an aged APPSWE/PS1ΔE9 murine model of Alzheimer’s disease
Minter, M. R., Hinterleitner, R., Meisel, M., Zhang, C., Leone, V., Zhang, X., Oyler-Castrillo, P., Zhang, X., Musch, M. W., Shen, X., et al. (2017) · Scientific Reports · doi:10.1038/s41598-017-11047-w · Cited by 292 · see citing works
Secreted protein lipocalin‐2 promotes microglial M1 polarization
Jang, E., Lee, S., Kim, J., Kim, J., Seo, J., Lee, W., Mori, K., Nakao, K., & Suk, K. (2013) · The FASEB Journal · doi:10.1096/fj.12-222257 · Cited by 198 · see citing works
Directed evolution of adeno-associated virus for efficient gene delivery to microglia
Lin, R., Zhou, Y., Yan, T., Wang, R., Li, H., Wu, Z., Zhang, X., Zhou, X., Zhao, F., Zhang, L., et al. (2022) · Nature Methods · doi:10.1038/s41592-022-01547-7 · Cited by 171 · see citing works
Nuclear Receptors License Phagocytosis by Trem2<sup>+</sup>Myeloid Cells in Mouse Models of Alzheimer's Disease
Savage, J. C., Jay, T., Goduni, E., Quigley, C., Mariani, M. M., Malm, T., Ransohoff, R. M., Lamb, B. T., & Landreth, G. E. (2015) · The Journal of Neuroscience · doi:10.1523/jneurosci.4586-14.2015 · Cited by 154 · see citing works
“Adult microglia/macrophages were isolated as described previously (Cardona et al., 2006).”
MethodSelf-citation
Integrated expression profiles of mRNA and miRNA in polarized primary murine microglia
Freilich, R. W., Woodbury, M. E., & Ikezu, T. (2013) · PLoS ONE · doi:10.1371/journal.pone.0079416 · Cited by 160 · see citing works
Microglia change from a reactive to an age-like phenotype with the time in culture
Caldeira, C., Oliveira, A. F., Cunha, C., Vaz, A. R., FalcÃ£o, A. S., Fernandes, A., & Brites, D. (2014) · Frontiers in Cellular Neuroscience · doi:10.3389/fncel.2014.00152 · Cited by 156 · see citing works
Analyzing microglial phenotypes across neuropathologies: a practical guide
Schwabenland, M., Brück, W., Priller, J., Stadelmann, C., Lassmann, H., & Prinz, M. (2021) · Acta Neuropathologica · doi:10.1007/s00401-021-02370-8 · Cited by 149 · see citing works
Microglial transglutaminase-2 drives myelination and myelin repair via GPR56/ADGRG1 in oligodendrocyte precursor cells
Giera, S., Luo, R., Ying, Y., Ackerman, S. D., Jeong, S.-J., Stoveken, H. M., Folts, C. J., Welsh, C. A., Tall, G. G., Stevens, B., et al. (2018) · eLife · doi:10.7554/elife.33385 · Cited by 145 · see citing works
Drug development for alzheimer’s disease: Microglia induced neuroinflammation as a target
Dong, Y., Li, X., Cheng, J., & Hou, L. (2019) · International Journal of Molecular Sciences · doi:10.3390/ijms20030558 · Cited by 136 · see citing works
Where are we? The anatomy of the murine cortical meninges revisited for intravital imaging, immunology, and clearance of waste from the brain
Coles, J. A., Myburgh, E., Brewer, J. M., & McMenamin, P. G. (2017) · Progress in Neurobiology · doi:10.1016/j.pneurobio.2017.05.002 · Cited by 123 · see citing works
Silencing of TREM2 exacerbates tau pathology, neurodegenerative changes, and spatial learning deficits in P301S tau transgenic mice
Jiang, T., Tan, L., Zhu, X.-C., Zhou, J.-S., Cao, L., Tan, M.-S., Wang, H.-F., Chen, Q., Zhang, Y.-D., & Yu, J.-T. (2015) · Neurobiology of Aging · doi:10.1016/j.neurobiolaging.2015.08.019 · Cited by 111 · see citing works
Telomere shortening reduces Alzheimer’s disease amyloid pathology in mice
Rolyan, H., Scheffold, A., Heinrich, A., Begus-Nahrmann, Y., Langkopf, B. H., Hölter, S. M., Vogt-Weisenhorn, D. M., Liss, B., Wurst, W., Lie, D. C., et al. (2011) · Brain · doi:10.1093/brain/awr133 · Cited by 106 · see citing works
Isolation of highly enriched primary human microglia for functional studies
Rustenhoven, J., Park, T. I.-H., Schweder, P., Scotter, J., Correia, J., Smith, A. M., Gibbons, H. M., Oldfield, R. L., Bergin, P. S., Mee, E. W., et al. (2016) · Scientific Reports · doi:10.1038/srep19371 · Cited by 99 · see citing works
Bone Marrow Transplantation Confers Modest Benefits in Mouse Models of Huntington's Disease
Kwan, W., Magnusson, A., Chou, A., Adame, A., Carson, M. J., Kohsaka, S., Masliah, E., Möller, T., Ransohoff, R., Tabrizi, S. J., et al. (2012) · The Journal of Neuroscience · doi:10.1523/jneurosci.4846-11.2012 · Cited by 75 · see citing works
Self-citation
Interleukin 4 modulates microglia homeostasis and attenuates the early slowly progressive phase of amyotrophic lateral sclerosis
Rossi, C., Cusimano, M., Zambito, M., Finardi, A., Capotondo, A., Garcia-Manteiga, J. M., Comi, G., Furlan, R., Martino, G., & Muzio, L. (2018) · Cell Death & Disease · doi:10.1038/s41419-018-0288-4 · Cited by 77 · see citing works
cTag-PAPERCLIP Reveals Alternative Polyadenylation Promotes Cell-Type Specific Protein Diversity and Shifts Araf Isoforms with Microglia Activation
Hwang, H.-W., Saito, Y., Park, C. Y., Blachère, N. E., Tajima, Y., Fak, J. J., Zucker-Scharff, I., & Darnell, R. B. (2017) · Neuron · doi:10.1016/j.neuron.2017.08.024 · Cited by 74 · see citing works
Scavenger Receptor-A deficiency impairs immune response of microglia and astrocytes potentiating Alzheimer’s disease pathophysiology
Cornejo, F., Vruwink, M., Metz, C., Muñoz, P., Salgado, N., Poblete, J., Andrés, M. E., Eugenín, J., & von Bernhardi, R. (2018) · Brain, Behavior, and Immunity · doi:10.1016/j.bbi.2017.12.007 · Cited by 59 · see citing works
Altered Neuroinflammation and behavior after traumatic brain injury in a mouse model of Alzheimer’s disease
Kokiko-Cochran, O., Ransohoff, L., Veenstra, M., Lee, S., Saber, M., Sikora, M., Teknipp, R., Xu, G., Bemiller, S., Wilson, G., et al. (2016) · Journal of Neurotrauma · doi:10.1089/neu.2015.3970 · Cited by 49 · see citing works
Self-citation
Glucocorticoid signaling in myeloid cells worsens acute CNS injury and inflammation
Sorrells, S. F., Caso, J. R., Munhoz, C. D., Hu, C. K., Tran, K. V., Miguel, Z. D., Chien, B. Y., & Sapolsky, R. M. (2013) · The Journal of Neuroscience · doi:10.1523/jneurosci.4705-12.2013 · Cited by 46 · see citing works
Mouse and human microglial phenotypes in Alzheimer’s disease are controlled by amyloid plaque phagocytosis through Hif1α
Grubman, A., Choo, X. Y., Chew, G., Ouyang, J. F., Sun, G., Croft, N. P., Rossello, F. J., Simmons, R., Buckberry, S., Landin, D. V., et al. (2019) · doi:10.1101/639054 · Cited by 7 · see citing works

Dispase rapidly and effectively purifies Schwann cells from newborn mice and adult rats.
Zhu, J., Qin, J., Shen, Z., et al. (2012) · PubMed · doi:10.3969/j.issn.1673-5374.2012.04.003 · Cited by 9 · see citing works
Preparation of single cells from pancreatic islets of adult rat by the use of dispase.
Ono, J., Takaki, R., & Fukuma, M. (1977) · Endocrinologia Japonica · doi:10.1507/endocrj1954.24.265 · Cited by 93 · see citing works
Dispase pretreatment enhances cell outgrowth from explanted human hair papillae.
Chiu, H., Chen, J., & Wu, Y.-J. (1993) · PubMed · Cited by 1 · see citing works
Alzheimer’s Disease Research Using Human Microglia
Lue, L., Beach, T. G., & Walker, D. G. (2019) · Cells · doi:10.3390/cells8080838 · Cited by 26 · see citing works
Differential Regional Vulnerability of the Brain to Mild Neuroinflammation Induced by Systemic LPS Treatment in Mice
Jung, H., Lee, H., Kim, D., Cheong, E., Hyun, Y., Yu, J., & Um, J. W. (2022) · Journal of Inflammation Research · doi:10.2147/jir.s362006 · Cited by 31 · see citing works

Authors: Cardona, A. E., Huang, D., Sasse, M. E., & Ransohoff, R. M.
Affiliations: Cleveland Clinic (US)
Corresponding author: Ransohoff, Richard M
ORCID: Cardona, Astrid E
Ransohoff, Richard M
Venue: Nature Protocols
Publisher: Springer Science and Business Media LLC
Published: 2006-11
Volume: 1
Issue: 4
Pages: 1947-1951
ISSN: 1750-2799 (ISSN-L)
1754-2189
DOI: 10.1038/nprot.2006.327
OpenAlex: W2088301466
Cited by: 236
References: 35
Citations by year: 2026: 3 · 2025: 7 · 2024: 5 · 2023: 8 · 2022: 20 · 2021: 24 · 2020: 15 · 2019: 11 · 2018: 20 · 2017: 17 · 2016: 15 · 2015: 12 · 2014: 19 · 2013: 17 · 2012: 11
FWCI: 1.32
Type: journal article
Language: en
Open access: closed
Licence: www.springer.com/tdm (tdm)
Locations: Nature Protocols (publishedVersion)
PubMed (publishedVersion)
Funders: National Multiple Sclerosis Society
National Institutes of Health
National Institute of Neurological Disorders and Stroke (NS32151, R01NS032151)
Topics: Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
Life Sciences › Neuroscience › Neurology › Neurological Disease Mechanisms and Treatments
Keywords: microglia isolation, microglial cells, mouse brain, collagenase digestion, Percoll gradient separation, infiltrating leukocytes, neuroinflammation, cell surface markers, adoptive transfer, RNA expression profiling
MeSH terms: Animals, Cell Separation, Flow Cytometry, RNA, Microglia, Mice

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review

Isolation of murine microglial cells for RNA analysis or flow cytometry

Abstract

Integrity signals

Claims the paper makes

Claims other papers make about this one

Cites

Cited by

Related

Record

About these checks