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. 2008 Oct 23:8:102.
doi: 10.1186/1471-213X-8-102.

Expression profiling of human glial precursors

James T Campanelli  1 , Robert W Sandrock, Will Wheatley, Haipeng Xue, Jianhua Zheng, Feng Liang, Jonathan D Chesnut, Ming Zhan, Mahendra S Rao, Ying Liu
Affiliations

Expression profiling of human glial precursors

James T Campanelli et al. BMC Dev Biol. .

Abstract

Background: We have generated gene expression databases for human glial precursors, neuronal precursors, astrocyte precursors and neural stem cells and focused on comparing the profile of glial precursors with that of other populations.

Results: A total of 14 samples were analyzed. Each population, previously distinguished from each other by immunocytochemical analysis of cell surface markers, expressed genes related to their key differentiation pathways. For the glial precursor cell population, we identified 458 genes that were uniquely expressed. Expression of a subset of these individual genes was validated by RT-PCR. We also report genes encoding cell surface markers that may be useful for identification and purification of human glial precursor populations.

Conclusion: We provide gene expression profile for human glial precursors. Our data suggest several signaling pathways that are important for proliferation and differentiation of human glial precursors. Such information may be utilized to further purify glial precursor populations, optimize media formulation, or study the effects of glial differentiation.

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Figures

Figure 1
Figure 1
Flow chart of generation of selected samples using MACS or double MACS in this work.
Figure 2
Figure 2
A2B5+/PSA-NCAM- MACS-sorted cells show properties of glial precursors. Immunocytochemistry analysis was done for the double MACS-enriched A2B5+/PSA-NCAM- (NAE) cells. Approximately 87% of NAE cells are A2B5+ (B) and ~9% are PSA-NCAM+ (D), They have high immunoreactivity to several glial (GFAP; C) and glial precursor markers (NG2 and PDGFRα; F and G). In contrast, lower percentage of cells show markers specific for the neuronal lineage, such as PSA-NCAM (D) and TuJ1 (βIII tubulin, E).
Figure 3
Figure 3
Scatter plots and dendrogram of related samples. Biological replicates of MACS sorted glial precursors (NAE) (A), and neurospheres (B), and technical replicates of FACS sorted glial precursors (HOPC) (C), and astrocyte precursors (hAPC) (D) show high R2 values indicating the array data are reliable. The dendrogram generated using BeadStudio software based on Euclidean distance (E) shows that replicates and samples with similar biological properties cluster closer to each other.
Figure 4
Figure 4
Unique genes that are expressed in glial precursors NAE. (A) shows the extent of overlap for expressed genes of three distinct types of cell populations, NAE, unsorted SM and astrocyte precursor hAPC. A total of 458 transcripts are expressed at ≥ 100 as detected by the array uniquely to NAE (detailed gene list can be found in Additional file 2). A selected group of genes (B) are validated by RT-PCR (C). In (B), the numbers in columns NAE, SM and APC represent the expression levels of these samples. The units are arbitrary as the signals are obtained by comparing with the background signals of the array chip. The numbers in columns NAE/SM and NAE/APC represent the ratios of the expression levels of the listed genes. The RT-PCR results (C) validate that all 10 genes listed in (B) have higher expression for NAE than for SM or APC, while the expression for the internal control GAPDH is similar across all three samples.
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