We have systematically optimized the concentrations of 20 components a previously published serum-free medium (Brewer and Cotman, Brain Res 494: 65-74, 1989) for survival rat embryonic hippocampal neurons after 4 days in culture. This supplement, B27, produced neuron above 60%, independent plating density 160 plated cells/mm2. For isolated cells (< 100 cells/mm2), at was still 45%, but could be rescued to 60% level 40 cells/mm2 by simply applying coverslip on top cells. suggests need...

Abstract Two fundamental questions about neuron cell culture were addressed. Can one serum‐free medium that was developed for optimum growth of hippocampal neurons support the from other regions brain? Is region specific state differentiation maintained in culture? To answer these questions, we isolated six rat brain regions, placed them B27/Neurobasal TM defined medium, and analyzed their morphology dependence on density after 4 days culture. Neuronal identity confirmed by immunostaining...

Abstract: The loss of ATP, which is needed for ionic homeostasis, an early event in the neurotoxicity glutamate and β‐amyloid (Aβ). We hypothesize that cells supplemented with precursor creatine make more phosphocreatine (PCr) create large energy reserves consequent neuroprotection against stressors. In serum‐free cultures, at 0.5‐1 m M was toxic to embryonic hippocampal neurons. Creatine &gt;0.1 greatly reduced toxicity. (1 ) could be added as late 2 h after achieve protection 24 h....

The brain depends on redox electrons from nicotinamide adenine dinucleotide (reduced form; NADH) to produce ATP and oxyradicals (reactive oxygen species [ROS]). Because ROS damage mitochondrial dysregulation are prominent in aging Alzheimer's disease (AD) their relationship the state is unclear, we wanted know whether an oxidative shift precedes these markers leads macromolecular a mouse model of AD. We used 3xTg-AD model, which displays cognitive deficits beginning at 4 months....

Summary Aging, a major risk factor in Alzheimer's disease ( AD ), is associated with an oxidative redox shift, decreased buffer protection, and increased free radical reactive oxygen species (ROS) generation, probably linked to mitochondrial dysfunction. While NADH the ultimate electron donor for many reactions, including phosphorylation, glutathione GSH ) ROS detoxifying cell. Here, we explored relative importance of neurodegeneration aging neurons from nontransgenic 3xTg‐ mice by...

For neurons to attach and remain in precise micropatterns for weeks culture, background molecules that nonpermissive extended culture durations need be identified. Nonpermissive of either polyethylene glycol (PEG) or the amino acid serine (C/sub 3/H/sub 7/NO/sub 3/) were evaluated. The foreground regions microstamped with 3-, 5-, 10-/spl mu/m lines poly-D-lysine (PDL), which promotes neural attachment growth. After 29 days compliance, fraction all live somata rested on desired PDL surface,...

Microcontact printing, facilitated by silane linker chemistry and high-relief stamps, creates precise patterns of proteins, which in turn control growth hippocampal neurons culture. This additive, multi-mask technique permits several different molecules to be patterned on the same substrate. The covalent technology relatively long-term (two-week) compliance stamped pattern against a polyethylene glycol background. When polylysine was adjacent laminin/polylysine mixture, neural somata...

Abstract Neuronal growth can be controlled in vitro by plating cells at low density and differential adhesion between the cell substrate. Primary cultures of rat hippocampal neurons were grown serum‐free culture on polylysine‐coated glass coverslips patterned selective laser ablation so as to leave grids polylysine with varying linewidths (3, 5, 10 μM), intersection distance (80, 120, 160 nodal (intersection) diameter (5, 10, 20 μM). Not only did somae strongly prefer unablated areas, but...

Technology has advanced to where it is possible design and grow - with predefined geometry surprisingly good fidelity living networks of neurons in culture dishes. Here we overview the elements design, emphasizing lithographic techniques that alter cell surface which turn influences attachment growth neural networks. Advanced capability this area makes desired complexity. Other issues addressed include influence glial cells media on activity potential for extending designs into three...

Age and Alzheimer's disease (AD) share some common features such as cognitive impairments, memory loss, metabolic disturbances, bioenergetic deficits, inflammation. Yet little is known on how systematic shifts in networks depend age AD. In this work, we investigated the global metabolomic alterations non-transgenic (NTg) triple-transgenic (3xTg-AD) mouse brain hippocampus a function of by using untargeted Ultrahigh Performance Liquid Chromatography-tandem Mass Spectroscopy (UPLC-MS/MS). We...