Product Name Human NeuronalBranching KitCatalog Number EP004Product Format 6,12, and 24 wellStorage 37°CGENERAL INFORMATION Our Human Neuronal Branching Kit possesses a cell body(soma), dendrites, and an axon. We do not use any Retinoic acid to help stimulate neuronalbranching in this model. Neuro Cells are cultured in our novel ECM Gel to help stimulateNeuronal Branching. End-user should start to see branch formation 48 hours after seedingthe Neuro cells.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 Human Neuronal Branching Kit contains all of the materials necessary to performmultiple assays in a 24-well format. The kit is designed that the testing materials.
compounds, conditioned media, or tissue explants, can be added into the system at anytime. The resulting effect on Neurites formation (tubular length, number of branches et al)can be monitored throughout the whole process under inverted fluorescence microscope.Reagents and Materials Provided:(1) 1 x vial of mixture of Human Brain Neuron ECs and RFP-tagged supporting cells (-80°Cor liquid 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 mesenchymal supporting cells).10. Gently remove condition medium. Be very careful not to damage the Neurites11. 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.