Use of Human Central Nervous System Cell Cultures in Neurotoxicity Testing

Sanfeliu, Coral; Cristòfol, Rosa; Torán, Núria; Rodrı́guez-Farré, Eduard; Kim, S.U.

doi:10.1016/s0887-2333(99)00065-x

Cited by 18 publications

(10 citation statements)

References 28 publications

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“…Cultures of trisomic neurons survived throughout the experiments but had more condensed cells than euploid ones, in agreement with an increase of apoptotic cells showed in a previous study (Sanfeliu et al ., 1999). However, trisomic astrocyte cultures were morphologically indistinguishable from normal astrocytes.…”

Section: Characterization Of Cultures Of Neurons and Astrocytes Euplsupporting

confidence: 91%

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Down's syndrome astrocytes have greater antioxidant capacity than euploid astrocytes

Sebastià

Cristòfol

Pertusa

et al. 2004

Eur J of Neuroscience

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Down's syndrome (trisomy 21) brain tissue is considered to be susceptible to oxidative injury, mainly because its increased Cu/Zn-superoxide dismutase (SOD1) activity is not followed by an adaptive rise in hydrogen peroxide metabolizing enzymes. In vitro, trisomic neurons suffer oxidative stress and degenerate. We studied the response of trisomy 21 neuron and astrocyte cultures to hydrogen peroxide injury and found that they were, respectively, more and less vulnerable than their euploid counterparts. Differences were detected 24 h after exposures in the region of 50 microm and 500 microm hydrogen peroxide for neuron and astrocyte cultures, respectively. Cytotoxicity results were paralleled by a decrease in cellular glutathione. In addition, trisomic astrocytes showed a lower basal content of superoxide ion and a higher clearance of hydrogen peroxide from the culture medium. In the presence of hydrogen peroxide, trisomic astrocytes maintained their concentration of intracellular superoxide and hydroperoxides at a lower level than euploid astrocytes. Consistent with these results, trisomic astrocytes in neuron coculture were more neuroprotective than euploid astrocytes against hydrogen peroxide injury. We suggest that SOD1 overexpression has beneficial effects on astrocytes, as it does in other systems with similarly high disposal of hydroperoxides. In addition to a higher enzymatic activity of SOD1, cultures of trisomic astrocytes showed slightly higher glutathione reductase activity than euploid cultures. Thus, trisomy 21 astrocytes showed a greater antioxidant capacity against hydrogen peroxide than euploid astrocytes, and they partially counteracted the oxidative vulnerability of trisomic neurons in culture.

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Section: Characterization Of Cultures Of Neurons and Astrocytes Euplsupporting

confidence: 91%

“…Astrocytes were seeded for experiments at 0.5 × 10 6 cells/mL (1.5 × 10 5 cells/cm 2 ) in multiwell plates, chamber slides or culture dishes, and used after 2–3 days. Cultures were characterized by immunocytochemistry procedures, as described elsewhere (Sanfeliu et al ., 1999).…”

Section: Cell Culturesmentioning

confidence: 99%

Down's syndrome astrocytes have greater antioxidant capacity than euploid astrocytes

Sebastià

Cristòfol

Pertusa

et al. 2004

Eur J of Neuroscience

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“…There were about 80% neurons at 8 and 14 DIV, and this percentage fell to 48% at 32 DIV and then further to 25% at 48 DIV. In a previous study (Sanfeliu et al,1999), the human cultures established at 8–14 DIV consisted of 75–85% neurons, 15–25% astrocytes, 1–2% microglia, and 0.1–1% oligodendrocytes, which is in agreement with the present results. Further immunostainings at 32 and 42 DIV showed that the content of microglia and oligodendrocytes remained below 2%.…”

Section: Resultssupporting

confidence: 93%

Spermine induces cell death in cultured human embryonic cerebral cortical neurons through N‐methyl‐D‐aspartate receptor activation

Vera

Martı́nez

Sanfeliu

2007

J of Neuroscience Research

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The polyamines putrescine, spermidine and spermine play important roles in cell proliferation, differentiation and the modulation of ion channel receptors. However, the function of increased concentrations of these compounds in brain injury and disease is unclear since they have been proposed as being both neuroprotective and neurotoxic. The effects of spermine and putrescine were studied in human primary cerebral cortical cultures containing both neurons and glia. No toxic effects were induced at 8 days in vitro (DIV) by either of the two polyamines at concentrations ranging from 0.3 µM to 2 mM. However, when the oxidative metabolisation of spermine that generates toxic by-products was induced by the presence of foetal calf serum, spermine caused cellular death with a LC 50 of approximately 50 μM. At 14 DIV, the coapplication of spermine 2 mM and glutamate 5 mM induced neuron cell death, but the effect of applying both components separately was null. Both spermine and glutamate were toxic to older neurons (26 to 42 DIV cultures), and here the coapplication of glutamate was found always to intensify the effect of spermine. Spermine showed greater toxicity than glutamate in neurons. Another effect observed is that glutamate, but not spermine, induced astrocyte swelling. Spermine toxicity was inhibited by both MK801 and ifenprodil, indicating a mechanism involving NMDA-receptor activation. Moreover, a strong spermine modulation of the NMDA receptor was demonstrated by the inhibition of glutamate toxicity by ifenprodil. Putrescine induced minor effects also as a neurotoxic agent. In conclusion, neuronal death by spermine can be induced by its toxic by-products as well as through NMDA-receptor action. The present results confirm the potentially harmful role of the polyamines in excitotoxicity-related human disorders.

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“…Their activation may interfere with neuronal cell health and integrity (Coyle and Schwarcz 2000;Moises et al 2002). Astroglial and microglial primary cultures have been used largely as models to evaluate their functions and their (Dawson et al 1994;Aschner 1998;Sanfeliu et al 1999;Streit et al 1999;Costa et al 2002;Vairano et al 2004). The flavonoid rutin and its aglycone quercetin have chemopreventive activity in a variety of colonic cancer cell lines and in animal models (Deschner et al 1991;Lipkin et al 1999;Yang et al 2000;Dihal et al 2006).…”

Section: Discussionmentioning

confidence: 99%

The flavonoid rutin induces astrocyte and microglia activation and regulates TNF-alpha and NO release in primary glial cell cultures

et al. 2007

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Astrocyte and microglia cells play an important role in the central nervous system (CNS). They react to various external aggressions by becoming reactive and releasing neurotrophic and/or neurotoxic factors. Rutin is a flavonoid found in many plants and has been shown to have some biological activities, but its direct effects on cells of the CNS have not been well studied. To investigate its potential effects on CNS glial cells, we used both astrocyte primary cultures and astrocyte/microglia mixed primary cell cultures derived from newborn rat cortical brain. The cultures were treated for 24 h with rutin (50 or 100 micromol/L) or vehicle (0.5% dimethyl sulfoxide). Mitochondrial function on glial cells was not evidenced by the MTT test. However, an increased lactate dehydrogenase activity was detected in the culture medium of both culture systems when treated with 100 micromol/L rutin, suggesting loss of cell membrane integrity. Astrocytes exposed to 50 micromol/L rutin became reactive as revealed by glial fibrillary acidic protein (GFAP) overexpression and showed a star-like phenotype revealed by Rosenfeld's staining. The number of activated microglia expressing OX-42 increased in the presence of rutin. A significant increase of nitric oxide (NO) was observed only in mixed cultures exposed to 100 micromol/L rutin. Enhanced TNFalpha release was observed in astrocyte primary cultures treated with 100 micromol/L rutin and in mixed primary cultures treated with 50 and 100 micromol/L, suggesting different sensitivity of both activated cell types. These results demonstrated that rutin affects astrocytes and microglial cells in culture and has the capacity to induce NO and TNFalpha production in these cells. Hence, the impact of these effects on neurons in vitro and in vivo needs to be studied.

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Use of Human Central Nervous System Cell Cultures in Neurotoxicity Testing

Cited by 18 publications

References 28 publications

Down's syndrome astrocytes have greater antioxidant capacity than euploid astrocytes

Down's syndrome astrocytes have greater antioxidant capacity than euploid astrocytes

Spermine induces cell death in cultured human embryonic cerebral cortical neurons through N‐methyl‐D‐aspartate receptor activation

The flavonoid rutin induces astrocyte and microglia activation and regulates TNF-alpha and NO release in primary glial cell cultures

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