Neuroscience 2004 Abstract
| Presentation Number: | 922.3 |
|---|---|
| Abstract Title: | Axon outgrowth in cryopreserved rat primary dorsal root ganglion neurons in culture. |
| Authors: |
Staines, W. A.*1,2
; Jacobsen, K.1
; Vaillancourt, K.3
; Di Ilio, Q.3
; Krantis, A.1,2
1Dept. of Cell. and Mol. Med., Univ. of Ottawa, Ottawa, Canada 2Centre for Res. in Biopharmaceutics and Biotech., Univ. of Ottawa, Ottawa, Canada 3ON, 451 Smyth Rd, K1H8M5, |
| Primary Theme and Topics |
Techniques in Neuroscience - Data analysis, physiological methods, statistics |
| Secondary Theme and Topics | Techniques in Neuroscience<br />- Mass spec and other biochemical and analytical methods |
| Session: |
922. Molecular, Anatomical, and Physiological Methods and Analysis Poster |
| Presentation Time: | Wednesday, October 27, 2004 10:00 AM-11:00 AM |
| Location: | San Diego Convention Center - Hall A-H, Board # GGG22 |
| Keywords: | AXON, PERIPHERAL NERVE, SENSORY NEURONS, DORSAL ROOT GANGLION |
As part of a series of feasibility studies on culture of cryopreserved dissociated neurons from varied regions of the nervous system we report on cryopreservation and subsequent functional studies on dorsal root ganglion cells and their axonal outgrowth. Rat embryonic dorsal root ganglion cells were isolated and dissociated using standard procedures. A method was developed to cryopreserve aliquots of 200 K DRG neurons which could be subsequently thawed and plated at 5 K per well (PDL coated 96 well plates) and grown in Neurobasal medium with B27. No further supplementation was required for survival to 19+ days in vitro. In culture, cryopreserved DRG neurons showed the same size and neurochemical type distribution as freshly dissociated dorsal root ganglion neurons. Over the first few days neuronal cell bodies formed ganglionated clusters and fasiculated axon bundles, but with the addition of mitotic inhibitors DRG neurons and their axons remain dispersed. Cells were transfected with eGFP using Transmessenger at 2 hrs to 18 hrs in vitro and outgrowth of fluorescent processes was monitored by recurrent digital imaging over the next 1 to 4 days. The cell soma, proximal and distal axon, growth cone and fine filopodia of transfected neurons were all readily imaged. This allowed characterization of dynamic parameters affecting axon outgowth and growth cone morphology.
Funded by: QBMCellScience.
Funded by: QBMCellScience.
<B>Conflict of Interest:</B> Conflict of Interest: Research funded by QBMCellScience, K. Vaillancourt and Q. Di Ilio are employees of QBMCellScience.
Sample Citation:
[Authors]. [Abstract Title]. Program No. XXX.XX. 2004 Neuroscience Meeting Planner. San Diego, CA: Society for Neuroscience, 2004. Online.
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