3D Human Neuronal Branching



Product Name 3D Human Neuronal Branching
Catalog Number EP011
Product Format 6,12, and 24 well
Storage 37°C

Our brains are full of fractals! In fact, they couldn’t function if not for fractal geometry. The
human brain comprises approximately 100 billion neurons. Amazingly, there are about 100
trillion synapses, or connections, among these brain cells. That’s an average of 1000
connections 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. The
axons reach out to make synaptic connections with the dendrites of other neurons. It is the
fractal branching pattern of the neuron’s axons and dendrites that allows them to
communicate with so many other cells. If neurons were shaped like cubes and neatly
packed 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 those
1) Demyelination
2) Axonal degeneration
3) Multiple sclerosis
4) Alzheimer’s disease
5) Nerve regeneration
6) HIV encephalitis
7) Parkinson’s disease
8) Neuromyelitis Optica
9) Myasthenia gravis
10)Charcot-Marie-Tooth disease

The 3D Human Neuronal Branching contains all of the materials necessary to perform
multiple angiogenesis assays in a 24-well format. The kit is designed that the testing
materials, i.e. compounds, conditioned media, or tissue explants, can be added into the
system at any time, ranging from the onset of advanced Neuronal Branching. The resulting
effect on tubule 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 Neurons and RFP-tagged supporting cells (-80°C or
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 1
1. Pre warm Endo-Growth Medium to 37ºC in a water bath
2. 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 2
9. Take the plate from the incubator and examine cells under inverted fluorescence
microscopy (Human Brain Neurons should sparsely and evenly distributed among RFP
positive human supporting cells).
10. Wash the cells one with 2 ml of PBS
11. Add 2.0ml of fresh Endo-Growth medium (control) or Experimental media (Endo-Growth
medium, 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 the
medium every 2 days until the end of the experiments.