We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Long-Term Primary Culture of Highly-Pure Rat Embryonic Hippocampal Neurons of Low-Density.
- Authors
Hao Yang; Rui Cong; Luo Na; Gong Ju; Si-Wei You
- Abstract
In order to develop a simplified method for long-term primary culture of highly-pure rat embryonic hippocampal neurons of low-density (103 cells/cm2), we optimized and modified conventional culturing methods. The modifications of our simplified method include: (1) combinational application of two growth substrates, tail collagen and poly-L-lysine, to coat plastic culture dishes and coverslips for a better neuronal attachment; (2) dissociation of hippocampal tissues with combinational use of two milder enzymes (collagenase and dispase) and trypsin of a lower concentration to minimize enzymatic damages to cultured neurons; (3) a cell pre-plating step to preliminarily eliminate the contaminating non-neuronal cells; (4) a modified culture medium as a critical step to promote highly pure neurons of low-density for a long term; and (5) appropriately reduced frequency and volume of refreshment of the culture medium. Using our modified method, the β-tubulin III-immunostained and Hoechst 33342 counterstained neurons harvested a steady and healthy growth with a longer culture time of over 35 days, and a clear distinction between TAU-1- and MAP2-immunoreactive neurites was apparent at the early culturing period. In addition, the purity of neurons was over 95% at the different time points in comparison with the control culture using conventional serum-free method in which most neurons degenerated and died within 5 days. Thus, our modified method proved to be a simple, feasible as well as time- and resource-saving approach for a long-term survival of pure rat embryonic hippocampal neurons of low-density.
- Publication
Neurochemical Research, 2010, Vol 35, Issue 9, p1333
- ISSN
0364-3190
- Publication type
Academic Journal
- DOI
10.1007/s11064-010-0189-0