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Product Name 3D Human Neuronal BranchingCatalog Number EP011Product Format 6,12, and 24 wellStorage 37°C
GENERAL INFORMATIONOur brains are full of fractals! In fact, they couldn’t function if not for fractal geometry. Thehuman brain comprises approximately 100 billion neurons. Amazingly, there are about 100trillion synapses, or connections, among these brain cells. That’s an average of 1000connections for a given cell, though some neurons may only make a single connection,while others may have hundreds of thousands of synapses with cells all over the brain. Theaxons reach out to make synaptic connections with the dendrites of other neurons. It is thefractal branching pattern of the neuron’s axons and dendrites that allows them tocommunicate with so many other cells. If neurons were shaped like cubes and neatlypacked into the brain, one neuron could only connect with at most 6 other cells.Our 3D Human Neuronal Branching possesses a cell body (soma), dendrites, and an axon.We do not use any Retinoic acid to help stimulate neuronal branching in this model.This model can be used to study the following disease, but not limited to thoseapplications:1) Demyelination2) Axonal degeneration3) Multiple sclerosis4) Alzheimer’s disease5) Nerve regeneration6) HIV encephalitis7) Parkinson’s disease8) Neuromyelitis Optica9) Myasthenia gravis10)Charcot-Marie-Tooth disease
The 3D Human Neuronal Branching contains all of the materials necessary to performmultiple angiogenesis assays in a 24-well format. The kit is designed that the testingmaterials, i.e. compounds, conditioned media, or tissue explants, can be added into thesystem at any time, ranging from the onset of advanced Neuronal Branching. The resultingeffect on tubule formation (tubular length, number of branches et al) can be monitoredthroughout the whole process under inverted fluorescence microscope.Reagents and Materials Provided:(1) 1 x vial of mixture of Human Brain Neurons and RFP-tagged supporting cells (-80°C orliquid N2)(2) 1 x 24-well Alpha Coat Solution coated plate (Room temperature, for 2 months)(3) 1 x 500ml of Endo-Growth Medium (4°C)Protocols: Day 11. Pre warm Endo-Growth Medium to 37ºC in a water bath2. Accurately pipette 24ml Endo-Growth Medium into a 50ml Falcon tube;3. Rapidly thaw the vial of cryopreserved cells in a 37ºC water bath;4. Transfer all cells gently into 24ml pre warmed Endo-growth medium;5. Mix well the cells gently using a serological pipette;6. Add 1.0ml of cell suspension to each well of the pre coated 24-well plate.7. Make sure the cells are evenly dispersed in the wells.8. Place the plate in an incubator (37ºC, 5% CO2 and humidified).Day 29. Take the plate from the incubator and examine cells under inverted fluorescencemicroscopy (Human Brain Neurons should sparsely and evenly distributed among RFPpositive human supporting cells).10. Wash the cells one with 2 ml of PBS11. Add 2.0ml of fresh Endo-Growth medium (control) or Experimental media (Endo-Growthmedium, plus pro- or anti-angiogenic regents according to customer’s needs).12. Place the plate back into the incubator. Day 4, 6, 8, 10, 12, and 14…… 13. Replace themedium every 2 days until the end of the experiments
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