PRIMARY NEURONAL AND ASTROCYTES CULTURES
Short term primary cultures from different parts of brain or spinal cord tissue are used for a variety of experimental conditions for basic and applied neuronal research. Therapeutic or experimental agents are tested for their potential use on cultured primary cells before launching in-vivo studies.
KTI has standardized protocols to generate neuron rich cultures, astrocyte rich cultures or mixed cultures of proportions relatively un-tampered by any treatments or supplements. We have taken over the burden of animal care & culture care so you do not have to, instead you can focus on crucial experimental and/or drug development and evaluation aspects. We are also tuned to modifying our protocol or using your protocol to serve your specific needs.
We prepare these cultures from 1-2 day postnatal pups or timed embryos of common laboratory animals such as rats and mice. We buy animals from reputed animal breeding colonies with a wide selection of strains and/or sub-species. Cultures can be provided ‘live’ within 40miles radius of our Beechcraft facility in Gaithersburg, MD in 60mm Petri-dishes, multi-well plates, chamber slides or culture-flasks in airtight container filled with 5% CO2. For longer distances, they can be provided ‘live’ in flasks or as cryo-preserved cells in vials on dry-ice. For specific needs, aliquots of complete culture medium are furnished along with the cultures.
Brief protocol to establish neuronal cultures adapted from Liu, Ginis et al, 2000; Am. J Physiol (Cell Physiol), ; 278 (1); C144) is outlined below.
- Dissect out the brain tissue from the animal in dissection medium.
- Remove meninges and remove all undesirable parts of the brain.
- Transfer the desired region of the brain (for example cortex and/or hippocampus) from which culture needs to be prepared. Clean out any meninges left behind.
- Trypsinize the selected brain region and gently triturate to release cells.
- Plate cells in poly-l-lysine coated plates in desired cell density in the desired neuronal or astrocyte specific medium.
- Change medium according to the specific needs of the culture.
