A reliable primary human CNS culture protocol for morphological studies of dendritic and synaptic elements.
Primary dissociated human fetal forebrain cultures were grown in defined serum-free conditions. At 4 weeks in vitro the cultures contained abundant morphologically well differentiated neurons with complex dendritic arbors. Astrocytic proliferation was negligible without the use of antimitotic agents. Confocal scanning laser microscopy (CSLM) and electron microscopy confirmed the presence of a dense neuropil, numerous cell-cell contacts and synapses. Neurons expressed a variety of proteins including growth associated protein-43 (GAP43), microtubule associated protein-2ab (MAP), class-III beta tubulin (C3BT), neurofilaments (NF), synaptophysin (SYN), parvalbumin (PA) and calbindin (CB). The cultures have proven to be reliable and simple to initiate and maintain for many weeks without passaging. They are useful in investigations of dendritic growth and injury of primary human CNS neurons.
Department of Pathology, London Health Sciences Centre, University of Western Ontario, London, Ont., Canada N6A 5C1. rhammond@julian.uwo.ca
Robert R
RR
Hammond
Sam
S
Iskander
Cristian L
CL
Achim
Stephen
S
Hearn
Jane
J
Nassif
Clayton A
CA
Wiley
D064026:Calbindins; D018929:Cell Culture Techniques / Q000379:methods; D003712:Dendrites / Q000648:ultrastructure*; D005333:Fetus; D019922:GAP-43 Protein / Q000096:biosynthesis; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis; D016900:Neurofilament Proteins / Q000096:biosynthesis; D009457:Neuroglia / Q000648:ultrastructure; D009474:Neurons / Q000378:metabolism / Q000648:ultrastructure*; D010320:Parvalbumins / Q000096:biosynthesis; D016548:Prosencephalon; D064030:S100 Calcium Binding Protein G / Q000096:biosynthesis; D013569:Synapses / Q000648:ultrastructure*; D016708:Synaptophysin / Q000096:biosynthesis; D014404:Tubulin / Q000096:biosynthesis
D064026:Calbindins; D019922:GAP-43 Protein; D008869:Microtubule-Associated Proteins; D016900:Neurofilament Proteins; D010320:Parvalbumins; D064030:S100 Calcium Binding Protein G; D016708:Synaptophysin; D014404:Tubulin