A new method to isolate microglia from adult mice and culture them for an extended period of time

Moussaud, Simon; Draheim, Henning J.

doi:10.1016/j.jneumeth.2010.01.017

Cited by 95 publications

(98 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, microglial activation was seen more as a 24 problem than an advantage, and a myriad of drugs have been 25 suggested to prevent microglial activation (Aisen, 2002;Block and 26 Hong, 2005; Gao et al, Rock and Peterson, 2006). However, 27 similarities between macrophages and microglia led to the 28 suspicion that the latter might also have an alternative activation 29 pathway that could be beneficial instead of harmful for neurons.…”

Section: Q5mentioning

confidence: 99%

Alternatively activated microglia and macrophages in the central nervous system

Franco

Fernández-Suárez

2015

Progress in Neurobiology

575

461

View full text Add to dashboard Cite

Section: Q5mentioning

confidence: 99%

Alternatively activated microglia and macrophages in the central nervous system

Franco

Fernández-Suárez

2015

Progress in Neurobiology

575

461

View full text Add to dashboard Cite

“…Percoll density gradient methods have been used to isolate microglial cells from adult mice [ 6,21,28 ] . Cardona et al, who recommended the use of fl ow cytometric analysis and PCR studies used a 70 %/40 % gradient, and the cells were observed at the interphase of 40 and 70 % [ 6 ] .…”

Section: Resultsmentioning

confidence: 99%

“…Recently, several groups have established microglial cell cultures from adult CNS using density gradient methods [ 6,21,28 ] . These microglial cells are adherent and showed morphological changes depending on the stimulant.…”

Section: Introductionmentioning

confidence: 99%

Establishment and Characterization of Primary Adult Microglial Culture in Mice

Ohtaki

Tsumuraya

Song

et al. 2013

Acta Neurochirurgica Supplement

View full text Add to dashboard Cite

Microglial cells account for approximately 12-15 % of the cells in the central nervous system (CNS). Microglial cells are polarized by pathological stimuli such as cytokines, chemokines, and growth factors, and play important roles in the deterioration and repair of the CNS. Here, we established cultures of primary microglial cells isolated from the brains of adult C57/BL6 mice using Percoll density gradients. The cells were cultured and stained with antibodies against CD11b, glial fibrillary acidic protein, myelin basic protein and NeuN to determine microglial, astroglial, oligodendroglial, and neuronal cells respectively. Moreover, the cells were exposed to interferon-γ (IFNγ) plus interleukin-1β (IL-1β) or IL-4 for 24 h to demonstrate the activating phenotypes with inducible nitric oxide synthase (iNOS), Ym1, and Iba-1 immunoblotting. At least 95 % of the cultured cells were CD11b-positive and -negative for astroglial, neuronal, and oligodendrocyte markers. IFNγ plus IL-1β treatment resulted in classical activation, which was represented by an increase in iNOS. The cells also displayed alternative activation, which increased Ym1 when treated with IL-4. The present study indicates that the microglial cells isolated as described here are a useful tool for elucidating adult microglial function.

show abstract

“…Microglia, a kind of glial cells derived from early embryonic myeloid-monocytic cells, are antigen presenting cells in the CNS (Cardona et al 2006;Moussaud and Draheim 2010). The major functions of microglia within the brain are keeping homeostasis and promoting inflammation in the infected or damaged tissue Mariani and Kielian 2009;Nelson et al 2002).…”

Section: Introductionmentioning

confidence: 99%

“…It is an important precondition to get large numbers of microglia efficiently and conveniently in vitro for further research. Several methods are employed to obtain microglia, including separation through gradient density centrifugation (Gingras et al 2007; Moussaud and Draheim 2010), fluorescence-activated or magnetic-activated cell sorting (Ford et al 1995;Marek et al 2008), and isolation by mild trypsinization (Saura et al 2003). Nevertheless, these methods are not easy to operate, costing much time and a larger population of animals are killed.…”

Section: Introductionmentioning

confidence: 99%

A Much Convenient and Economical Method to Harvest a Great Number of Microglia

Qin

Tian

et al. 2011

Cell Mol Neurobiol

View full text Add to dashboard Cite

Microglia, implicating in such neuro-pathologies as brain inflammation, neurodegeneration, glioma, and neurogenesis, play an important role in central nervous system. Advanced research on microglia is crucial in exploring the neuro-pathology and neuro-physiology of these diseases, so how to culture large numbers of microglia in vitro becomes the base of a research. The wildly used method, at present, obtaining microglia from murine cannot fulfill the requirement of research, costing too much time and needing too many rats. We intend to introduce an optimized method that can harvest large quantities of microglia with high purity. Neonatal 2-3 days old Wistar rats were sacrificed and the cerebral cortices were trypsinized. We primarily cultured mixed cortical cells for 8-10 days. The microglia were harvested from the liquid supernatant; the left cells in the mixed cortical glial culture were passaged at a 1:2 density. After another 8-10 days of culture, microglia were collected again. And then, we passaged the left cells again for acquiring microglia from the third collection. We did not add additional mitogens in the experiment. At last, on average, 7.0 × 10(6) microglia were collected from one neonatal rat. By this modified method, much more microglia can be effectively and easily harvested comparing with the usual protocol before. We compared the characteristics of microglia harvested from these three passages, such as morphology, phenotype, purity, and abilities on proliferation, secretion, and phagocytosis. The cells presented typical microglia morphology, having phenotype markers of CD11b/c and CD45. The microglia from these three passages retained similar phagocytosis and secretion functions. Expanded population of microglia for investigation can be provided by this easy method in a short time with little cost and few rats.

show abstract

A new method to isolate microglia from adult mice and culture them for an extended period of time

Cited by 95 publications

References 49 publications

Alternatively activated microglia and macrophages in the central nervous system

Alternatively activated microglia and macrophages in the central nervous system

Establishment and Characterization of Primary Adult Microglial Culture in Mice

A Much Convenient and Economical Method to Harvest a Great Number of Microglia

Contact Info

Product

Resources

About