Cell death and proliferation in acute slices and organotypic cultures of mammalian CNS

Lossi, Laura; Alasia, Silvia; Salio, Chiara; Merighi, Adalberto

doi:10.1016/j.pneurobio.2009.01.002

Cited by 127 publications

(134 citation statements)

References 454 publications

(280 reference statements)

Supporting

Mentioning

125

Contrasting

Unclassified

Order By: Relevance

“…Using a slice of brain tissue, as in the model proposed here, overcomes the disadvantages described above and has the added advantage of being used in conjunction with electrophysiological techniques (Gahwiler et al, 1997;Lossi et al, 2009). In this way, we were able to measure, in real-time, changes in membrane potential (i.e.…”

Section: Discussionmentioning

confidence: 99%

“…Consequently, neurons in culture lack normal synaptic contacts and interactions with other neurons as well as with other cerebral cell types (i.e. glial cells; Lossi et al, 2009). The in vitro brain slice model may be lower throughput than the cell culture model, however, it is far more physiologically accurate.…”

Section: E-mail Address: Tsaleh@upeica (Tm Saleh)mentioning

confidence: 99%

“…The in vitro brain slice model may be lower throughput than the cell culture model, however, it is far more physiologically accurate. Within each slice, cytoarchitecture is maintained and thus many of the cell-to-cell interactions and neuronal networks remain intact (Gahwiler et al, 1997;Noraberg et al, 2005;Lossi et al, 2009). Hence, this model is well suited for physiological experiments to assess mechanism of action of drugs as well as to study neurophysiological changes that occur with stroke.…”

Section: E-mail Address: Tsaleh@upeica (Tm Saleh)mentioning

confidence: 99%

See 2 more Smart Citations

A novel method for inducing focal ischemia in vitro

Richard

Saleh

Bahh³

et al. 2010

Journal of Neuroscience Methods

View full text Add to dashboard Cite

Current in vitro models of stroke involve applying oxygen-glucose deprived (OGD) media over an entire brain slice or plate of cultured neurons. Thus, these models fail to mimic the focal nature of stroke as observed clinically and with in vivo rodent models of stroke. Our aim was to develop a novel in vitro brain slice model of stroke that would mimic focal ischemia and thus allow for the investigation of events occurring in the penumbra. This was accomplished by focally applying OGD medium to a small portion of a brain slice while bathing the remainder of the slice with normal oxygenated media. This technique produced a focal infarct on the brain slice that increased as a function of time. Electrophysiological recordings made within the flow of the OGD solution ("core") revealed that neurons rapidly depolarized (anoxic depolarization; AD) in a manner similar to that observed in other stroke models. Edaravone, a known neuroprotectant, significantly delayed this onset of AD. Electrophysiological recordings made outside the flow of the OGD solution ("penumbra") revealed that neurons within this region progressively depolarized throughout the 75 min of OGD application. Edaravone attenuated this depolarization and doubled neuronal survival. Finally, synaptic transmission in the penumbra was abolished within 50 min of focal OGD application. These results suggest that this in vitro model mimics events that occur during focal ischemia in vivo and can be used to determine the efficacy of therapeutics that target neuronal survival in the core and/or penumbra.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: E-mail Address: Tsaleh@upeica (Tm Saleh)mentioning

confidence: 99%

Section: E-mail Address: Tsaleh@upeica (Tm Saleh)mentioning

confidence: 99%

See 1 more Smart Citation

A novel method for inducing focal ischemia in vitro

Richard

Saleh

Bahh³

et al. 2010

Journal of Neuroscience Methods

View full text Add to dashboard Cite

show abstract

“…Slicing is performed with a vibrating microtome at high amplitude and very slow speed. Slice thickness varies according to specimen and the type of experiment, from 150 to 400 μm (Lossi et al, 2009). Over the years, slice culture systems have been successfully established from a variety of brain regions including hippocampus (far more frequently), striatum, cortex, spinal cord and cerebellum.…”

Section: Preparation Of Organotypic Cultures Of Cns Tissuementioning

confidence: 99%

“…Moreover, when organotypic slices are exposed to certain toxic conditions (oxygen-glucose deprivation, neurotoxins, glutamate-mediated excitotoxicity) they develop many pathophysiological features found in brain disorders and consequently, brain organotypic slices can be used as ex-vivo models of CNS diseases. Nowadays, organotypic models for global cerebral ischemia, ischemic stroke, Alzheimer's disease, PD, HD, TBI, epilepisa and amyotrophic lateral sclerosis have been described (For review see (Noraberg et al, 2005;Sundstrom et al, 2005;Cho et al, 2007;Cimarosti & Henley, 2008;Lossi et al, 2009)). Among them, hippocampal organotypic slices exposed to oxygen-glucose deprivation, a model for global cerebral ischemia is the most commonly used.…”

Section: Preparation Of Organotypic Cultures Of Cns Tissuementioning

confidence: 99%

Tissue Engineering for Tissue and Organ Regeneration

Eberli¹

2011

View full text Add to dashboard Cite

Organotypic brain explant culture as a drug evaluation system for malignant brain tumors

et al. 2017

View full text Add to dashboard Cite

Therapeutic options for malignant brain tumors are limited, with new drugs being continuously evaluated. Organotypic brain slice culture has been adopted for neuroscience studies as a system that preserves brain architecture, cellular function, and the vascular network. However, the suitability of brain explants for anticancer drug evaluation has been unclear. We here adopted a mouse model of malignant glioma based on expression of H‐RasV12 in Ink4a/Arf −/− neural stem/progenitor cells to establish tumor‐bearing brain explants from adult mice. We treated the slices with cisplatin, temozolomide, paclitaxel, or tranilast and investigated the minimal assays required to assess drug effects. Serial fluorescence‐based tumor imaging was sufficient for evaluation of cisplatin, a drug with a pronounced cytotoxic action, whereas immunostaining of cleaved caspase 3 (a marker of apoptosis) and of Ki67 (a marker of cell proliferation) was necessary for the assessment of temozolomide action and immunostaining for phosphorylated histone H3 (a marker of mitosis) allowed visualization of paclitaxel‐specific effects. Staining for cleaved caspase 3 was also informative in the assessment of drug toxicity for normal brain tissue. Incubation of explants with fluorescently labeled antibodies to CD31 allowed real‐time imaging of the microvascular network and complemented time‐lapse imaging of tumor cell invasion into surrounding tissue. Our results suggest that a combination of fluorescence imaging and immunohistological staining allows a unified assessment of the effects of various classes of drug on the survival, proliferation, and invasion of glioma cells, and that organotypic brain slice culture is therefore a useful tool for evaluation of antiglioma drugs.

show abstract

Cell death and proliferation in acute slices and organotypic cultures of mammalian CNS

Cited by 127 publications

References 454 publications

A novel method for inducing focal ischemia in vitro

A novel method for inducing focal ischemia in vitro

Tissue Engineering for Tissue and Organ Regeneration

Organotypic brain explant culture as a drug evaluation system for malignant brain tumors

Contact Info

Product

Resources

About