Document Detail


In vitro isolation of neural precursor cells from the adult pig subventricular zone.
MedLine Citation:
PMID:  19524610     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In order to improve cell therapy techniques, we have characterized a multipotent neural precursor cell isolation technique from the subventricular zone of adult pig brain. The pig is a non-primate species that is immunologically closest to human. The proliferative zone of this neurogenic structure was first localized in situ in the pig brain by Ki-67 immunohistochemistry, as a ventral subfield of the Nissl-stained subventricular zone. For in vitro cultures, the striatal forebrain was sampled from deeply anaesthetized adult pigs and SVZ tissue explants were immediately microdissected out and dissociated in the appropriate medium. Primary cell culture in the presence of EGF and bFGF allowed growth of spherical masses that exhibited sustained growth and self-renewal capacity through six subsequent passages. Molecular characterization using reverse transcription-polymerase chain reaction (RT-PCR) showed that expanded pro-differentiating neurospheres expressed markers of proliferation, neural stem cells, and committed neural progenitors. After growth factor removal, the spheres became adherent and were shown to contain the three neural cell lineages by triple immunocytofluorescence and confocal microscopy. The present protocol therefore allowed for in vitro expansion of pig brain primary cells that display capacities for proliferation, self-renewal, and multipotency, i.e., the cardinal features of multipotent neural precursor cells.
Authors:
Olivier Liard; Stéphanie Segura; Aurélie Pascual; Pierrette Gaudreau; Thierry Fusai; Emmanuel Moyse
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-06-12
Journal Detail:
Title:  Journal of neuroscience methods     Volume:  182     ISSN:  1872-678X     ISO Abbreviation:  J. Neurosci. Methods     Publication Date:  2009 Sep 
Date Detail:
Created Date:  2009-08-14     Completed Date:  2009-10-23     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7905558     Medline TA:  J Neurosci Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  172-9     Citation Subset:  IM    
Affiliation:
Service de Chirurgie Maxillo-faciale et Plastique de face, HIA Laveran, BP 50, 13998 Marseille Armées, France.
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MeSH Terms
Descriptor/Qualifier:
Animals
Biological Markers / analysis
Cell Proliferation
Cell Separation / methods*
Epidermal Growth Factor / metabolism
Fibroblast Growth Factor 1 / metabolism
Immunohistochemistry
Ki-67 Antigen / chemistry
Multipotent Stem Cells / physiology*
RNA, Messenger / biosynthesis,  genetics
Reverse Transcriptase Polymerase Chain Reaction
Swine
Telencephalon / cytology*
Tissue Fixation
Chemical
Reg. No./Substance:
0/Biological Markers; 0/Ki-67 Antigen; 0/RNA, Messenger; 104781-85-3/Fibroblast Growth Factor 1; 62229-50-9/Epidermal Growth Factor

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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