Human medulloblastoma cell line DEV is a potent tool to screen for factors influencing differentiation of neural stem cells

Abstract

The aim of our study was to investigate whether a human neural cell line could be used as a reliable screening tool to examine the functional conservation, in humans, of transcription factors involved in neuronal or glial specification in other species. Gain-of-function experiments were performed on DEV cells, a cell line derived from a human medulloblastoma. Genes encoding nine different transcription factors were tested for their influence on the process of specification of human DEV cells towards a neuronal or glial fate. In a first series of experiments, DEV cells were transfected with murine genes encoding transcription factors known to be involved in the neuronal differentiation cascade. Neurogenins-1, -2, and -3; Mash-1; and NeuroD increased the differentiation of DEV cells towards a neuronal phenotype by a factor of 2-3.5. In a second series of experiments, we tested transcription factors involved in invertebrate glial specification. In the embryonic Drosophila CNS, the development of most glial cells depends on the master regulatory gene glial cell missing (gcm). Expression of gcm in DEV cells induced a twofold increase of astrocytic and a sixfold increase of oligodendroglial cell types. Interestingly, expression of tramtrack69, which is required in all Drosophila glial cells, resulted in a fourfold increase of only the oligodendrocyte phenotype. Expression of the related tramtrack88 protein, which is not expressed in the fly glia, or the C. elegans lin26 protein showed no effect. These results show that the Drosophila transcription factor genes tested can conserve their function upon transfection into the human DEV cells, qualifying this cell line as a screening tool to analyze the mechanisms of neuronal and glial specification. J. Neurosci. Res. 65:17-23, 2001.

Cancer stem cells in nervous system tumors

“…Microarray analysis of human medulloblastomas suggests a similarity of gene expression with normal developing brain cells (Pomeroy et al, 2002). In a recent study, transfection of a medulloblastoma cell line with transcription factors specified a factor-dependent adoption of either neuronal or glial cell fates (Buzanska et al, 2001). Finally, mutations that dysregulate the pathways that control normal stem cell self-renewal (such as the sonic hedgehog pathway) cause brain tumors, emphasizing the mechanistic similarities between normal stem cell and cancer cell self-renewal (Dahmane et al, 2001;Ruiz i Altaba et al, 2002;Pardal et al, 2003).…”

A novel, neural potential of non-hematopoietic human umbilical cord blood stem cells

“…Our special interest focused on transcriptional regulation of glial or neuronal fate decision undertaken by non-differentiated human neural stem-like cells. For that purpose the cells were transfected with evolutionary conserved murine or invertebrate genes encoding transcription factors known to be involved in the neuronal or glial differentiation cascade (Buzanska et al, 2001b (Singh et al, Oncogene, 2004).…”

Glial differentiation and the Gcm pathway

“…The DNA-binding domain represents the structural hallmark of all gcm genes and the only region of significant homology among the different Gcm family members (Akiyama et al, 1996). Moreover, introduction of the fly gcm gene into a medulloblastoma line as well as into HeLa cells induces the expression of a glial marker (Buzanska et al, 2001;Soustelle et al, 2007). Moreover, introduction of the fly gcm gene into a medulloblastoma line as well as into HeLa cells induces the expression of a glial marker (Buzanska et al, 2001;Soustelle et al, 2007).…”