Comparative Analysis of the Frequency and Distribution of Stem and Progenitor Cells in the Adult Mouse Brain

Abstract

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 m 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 neurosphereforming cells detected in individual 400 m 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. STEM CELLS 2008;26:979 -987 Disclosure of potential conflicts of interest is found at the end of this article.

Oxygen Tension Within the Neurogenic Niche Regulates Dopaminergic Neurogenesis in the Developing Midbrain

“…However, further studies are warranted to investigate whether the observed changes in dopaminergic neurogenesis translate into functional changes of the dopaminergic system in the postnatal and/or adult brain. Detailed knowledge of the composition of the midbrain neurogenic niche harboring dopaminergic NPC capacity will eventually help to develop restorative strategies for neurodegenerative diseases such as Parkinson's disease by both recruitment of endogenous progenitor cells from the niche [50][51][52] and improvement of dopaminergic cell graft. Our study, therefore, provides the initial framework for future studies on the molecular mechanisms mediating oxygen regulation of dopaminergic neurogenesis within the midbrain neurogenesis as its natural environment.…”

Stem cells of the adult mammalian brain and their niche

“…66,2009 Review Article tional data are still lacking [142]. In addition, markers of proliferation such as proliferating cell nuclear antigen (PCNA) and Ki67 are frequently used to label cells within the cell cycle; however, their association with NSCs has not been demonstrated, partially due to the lack of markers.…”

Multivalent epigenetic marks confer microenvironment-responsive epigenetic plasticity to ovarian cancer cells

“…Table S2). While other in vitro cultured multiaggregate spheroids were enriched in stem cells (i.e., less differentiated), this property would appear to be tissue-specific (including neural and mammary tissues), 63,64 as other sphere-forming cells actually demonstrate reduced differentiation capacity (i.e., reduced plasticity/stemness). 65,66 It is conceivable that our in vitro spheroids resemble those in the abovementioned melanoma differentiation studies.…”