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TISSUE-SPECIFIC STEM CELLS |
aQueensland Brain Institute, The University of Queensland, Brisbane, QLD 4072, Australia,
bSchool of Medicine, Isfahan University of Medical Sciences, Isfahan 81744-176, Iran,
cSchool of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran,
dPrince of Wales Medical Research Institute, Randwick, NSW 2031, Australia,
eDepartment of Psychology, McGill University, Montreal, Quebec, Canada H3A 1B1
Key Words. Neural stem cell • Precursor • Bromodeoxyuridine • Label-retaining cell • Neurosphere
Correspondence: Correspondence: Rodney L. Rietze, M.Sc., Ph.D., Queensland Brain Institute, The University of Queensland, Brisbane QLD 4072, Australia. Telephone: +61-7-3346-6351; Fax: +61-7-3346-6051; e-mail: rietze{at}uq.edu.au
Received on November 2, 2007;
accepted for publication on January 9, 2008.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLS EXPRESS January 17, 2008.
The neurosphere assay can detect and expand neural stem cells (NSCs) and progenitor cells, but it cannot discriminate between these two populations. Given two assays have purported to overcome this shortfall, we performed a comparative analysis of the distribution and frequency of NSCs and progenitor cells detected in 400 µm coronal segments along the ventricular neuraxis of the adult mouse brain using the neurosphere assay, the neural colony forming cell assay (N-CFCA), and label-retaining cell (LRC) approach. We observed a large variation in the number of progenitor/stem cells detected in serial sections along the neuraxis, with the number of neurosphere-forming cells detected in individual 400 µm sections varying from a minimum of eight to a maximum of 891 depending upon the rostral-caudal coordinate assayed. Moreover, the greatest variability occurred in the rostral portion of the lateral ventricles, thereby explaining the large variation in neurosphere frequency previously reported. Whereas the overall number of neurospheres (3730 ± 276) or colonies (4275 ± 124) we detected along the neuraxis did not differ significantly, LRC numbers were significantly reduced (1186 ± 188, 7 month chase) in comparison to both total colonies and neurospheres. Moreover, approximately two orders of magnitude fewer NSC-derived colonies (50 ± 10) were detected using the N-CFCA as compared to LRCs. Given only 5% of the LRCs are cycling (BrdU+/Ki-67+) or competent to divide (BrdU+/Mcm-2+), and proliferate upon transfer to culture, it is unclear whether this technique selectively detects endogenous NSCs. Overall, caution should be taken with the interpretation and employment of all these techniques.
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