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  • 1.
    Sievertzon, Maria
    et al.
    KTH, School of Biotechnology (BIO), Centres, KTH Genome Center.
    Wirta, Valtteri
    KTH, School of Biotechnology (BIO), Centres, KTH Genome Center.
    Mercer, Alex
    Meletis, Konstantinos
    Erlandsson, Rikard
    Wikström, Lilian
    Frisén, Jonas
    Lundeberg, Joakim
    KTH, School of Biotechnology (BIO), Centres, KTH Genome Center.
    Transcriptome analysis in primary neural stem cells using a tag cDNA amplification method2005In: BMC neuroscience (Online), ISSN 1471-2202, E-ISSN 1471-2202, Vol. 6, no 28, p. 13-Article in journal (Refereed)
    Abstract [en]

    Background: Neural stem cells ( NSCs) can be isolated from the adult mammalian brain and expanded in culture, in the form of cellular aggregates called neurospheres. Neurospheres provide an in vitro model for studying NSC behaviour and give information on the factors and mechanisms that govern their proliferation and differentiation. They are also a promising source for cell replacement therapies of the central nervous system. Neurospheres are complex structures consisting of several cell types of varying degrees of differentiation. One way of characterising neurospheres is to analyse their gene expression profiles. The value of such studies is however uncertain since they are heterogeneous structures and different populations of neurospheres may vary significantly in their gene expression.

    Results: To address this issue, we have used cDNA microarrays and a recently reported tag cDNA amplification method to analyse the gene expression profiles of neurospheres originating from separate isolations of the lateral ventricle wall of adult mice and passaged to varying degrees. Separate isolations as well as consecutive passages yield a high variability in gene expression while parallel cultures yield the lowest variability.

    Conclusions: We demonstrate a low technical amplification variability using the employed amplification strategy and conclude that neurospheres from the same isolation and passage are sufficiently similar to be used for comparative gene expression analysis.

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