Brain at cellular level

 The brain is composed of two broad classes of

cells: neurons and glia.

100 billion neurons in brain and about 1-5

trillion glial cells (that's 10-50 times more!)
.

 Neurons are electrically excitable cells in the

nervous system that process and transmit
information where as glia provide structural
and metabolic support, insulation, and
guidance of development
Preparation of cell suspensions
Transfer the minced tissue into centrifuge tubes with 10ml of Solution I

Centrifuge at 1000 rpm for 5-10 min

Resuspend the cell pellet in 10 ml of solution II,

Trypsinise in solution II by incubation at 37°C for 15 min with shaking

Add solution III containing trypsin inhibitor (10%FBS) to the suspension obtained
and centrifuge at 1500 rpm for 10 min.

• Discard the supernatant and suspend the cell pellet in 2ml of Solution III.

• Triturate the cell suspension with pipette (25 strokes).

Allow to stand for 5-10 min.

Aspirate and collect in DMEM media leaving the undissociated cell mass at the
bottom.

Centrifuge at 1500 rpm for 10 min.

Wash the pellet obtained with 2 ml of complete medium to remove traces

 Seeding and maintenance
Count the number of cells obtained in the cell suspension in the using Trypan blue
dye (1:10 dilution) using haemocytometer.

•Seed the cells after counting at a density of 2.7-3.0X 105 cells/ml of complete
medium in various coated Petri dishes.

•Incubate at 37°C in humidified 5% CO2 in a CO2 incubator