Prolonged cultivation of hippocampal neural precursor cells shifts their differentiation potential and selects for aneuploid cells

Abstract

Neural precursor cells (NPCs) are lineage-restricted neural stem cells with limited self-renewal, giving rise to a broad range of neural cell types such as neurons, astrocytes, and oligodendrocytes. Despite this developmental potential, the differentiation capacity of NPCs has been controversially discussed concerning the trespassing lineage boundaries, for instance resulting in hematopoietic competence. Assessing their in vitro plasticity, we isolated nestin+/Sox2+, NPCs from the adult murine hippocampus. In vitro-expanded adult NPCs were able to form neurospheres, self-renew, and differentiate into neuronal, astrocytic, and oligodendrocytic cells. Although NPCs cultivated in early passage efficiently gave rise to neuronal cells in a directed differentiation assay, extensively cultivated NPCs revealed reduced potential for ectodermal differentiation. We further observed successful differentiation of long-term cultured NPCs into osteogenic and adipogenic cell types, suggesting that NPCs underwent a fate switch during culture. NPCs cultivated for more than 12 passages were aneuploid (abnormal chromosome numbers such as 70 chromosomes). Furthermore, they showed growth factor-independent proliferation, a hallmark of tumorigenic transformation. In conclusion, our findings substantiate the lineage restriction of NPCs from adult mammalian hippocampus. Prolonged cultivation results, however, in enhanced differentiation potential, which may be attributed to transformation events leading to aneuploid cells.

Large-scale generation of human iPSC-derived neural stem cells/early neural progenitor cells and their neuronal differentiation

“…We do not recommend expanding NSCs/NPCs for more than 12 passages to reduce the risk of chromosomal aberration. 28 Freezing and thawing procedures of NSCs/ eNPCs are described in Supplementary Information.…”

Development of a cell line from the American eel brain expressing endothelial cell properties

“…This is suggested by recent research on the mammalian brain. In the human brain, neural progenitor cells are frequently found to be aneuploid (Rehen et al 2005;Devalle et al 2012), and during the long-term cultivation of mouse neural precursor cells, aneuploidy and an increased differentiation potential developed (Nguyen et al 2013).…”

“…Over the last decade, rodent and human NSCs/NPCs have been found to differentiate beyond neural lineage cells, such as neurons and astrocytes, and into other lineages, including endothelial cells (Wurmser et al 2004;Ii et al 2009;Sekiguchi et al 2013). During long-term culture, murine NPCs showed a dramatic reduction in their neuronal and glial differentiation capacity and an increase in their capacity for mesodermal differentiation (Nguyen et al 2013). Possibly, early-passage eelB cells can be used to explore the signals that cause neural stem cells to differentiate into specific cell types.…”

Stem Cells: How We Could Restore the Brain Function After Ischemic Damage

“…In this case, the source availability is also the main limiting factor, which is further complicated by the difficulty to isolate and successfully maintain these cells in vitro [18][19][20]. Moreover, it is unclear what would constitute the appropriate donor.…”