In vitro characterization of a murine oligodendrocyte precursor cell line (BO‐1) following spontaneous immortalization

Abstract

The understanding of oligodendrocyte differentiation is crucial for designing therapies of demyelinating diseases. Oligodendrocyte precursor cells are of particular interest in this context, because of their remyelinating potential. Permanent cell lines, which are a versatile tool for studying oligodendrocyte physiology, have been so far mainly established from the rat CNS. In the present study, we describe a novel murine oligodendrocyte precursor cell line (BO-1) established by spontaneous immortalization using light microscopy, immunocytochemical phenotyping and genetic analysis. BO-1 cells displayed a bi- to multipolar morphology and expressed early oligodendrocytic lineage markers, such as A2B5 and NG-2. Expression of pre-oligodendrocyte (O4, CNPase) and mature oligodendrocyte markers (e.g. myelin basic protein) was found in about 30% and 1.5% of the cells, respectively. Addition of serum, known to promote type-2 astrocyte differentiation, significantly increased the number of GFAP-positive cells, while thyroid hormones, (T3/T4) known to foster oligodendrocyte differentiation, did not substantially alter the antigenic and gene expression of myelin markers. This deficiency might be related to the high intrinsic proliferation rate of BO-1 cells that was unaltered upon removal of mitogenic factors. Expression of O4 and CNPase in BO-1 cells could be significantly increased by co-culture with primary astrocytes suggesting that the differentiating potential of BO-1 cells was influenced by environmental factors and may have to be fully explored in future studies. In summary, the novel murine BO-1 cell line shares several characteristics with oligodendrocyte precursor cells but displays a restricted differentiation into mature oligodendrocytes.

Phenotype, differentiation, and function differ in rat and mouse neocortical astrocytes cultured under the same conditions

“…Similarly, using a rat-adapted protocol, mouse oligodendrocyte precursor cells are not only more difficult to isolate and to maintain in culture for long time periods than rat cells are (Chen et al, 2007), but also substantially differ in the expression of classical oligodendrocyte markers (Fanarraga et al, 1995). To overcome this problem, Pringproa et al (2008) established a mouse oligodendrocyte precursor cell line (BO-1).…”

Isolation of cortical mouse oligodendrocyte precursor cells

“…Numerous oligodendroglial cell lines have been developed for the study of OL function such as the CG-4 permanent cell line and Oli-neu immortalized cell lines (Jung et al, 1995; Lin et al, 2006; Louis et al, 1992; Mock et al, 2006; Pringproa et al, 2008). However, there is considerable variability between these cell lines largely depending on the source (i.e.…”

“…chemically induced rodent tumors or spontaneous human tumors), isolation method, and growth conditions. Oftentimes, these lines maintain considerable proliferative potential, even once engrafted (a hallmark of their potential tumorigenicity), and may prove difficult for induction of differentiation (Lin et al, 2006; Pringproa et al, 2008). OPC cultures derived from the formation of neurospheres or oligospheres yield similar variability in genetic and phenotypic properties resulting in considerable departure from their in vivo counterparts (Chen et al, 2007; Zhao et al, 2010).…”

Defining the morphological phenotype: 2′,3′-cyclic nucleotide 3′-phosphodiesterase (CNPase) is a novel marker for in situ detection of canine but not rat olfactory ensheathing cells

“…CNPase is a well-established marker of the oligodendroglial lineage being expressed from early oligodendrocyte precursor cells (OPCs) to mature oligodendrocytes (Pringproa et al 2008). However, there have been controversial reports regarding the expression in OECs and Schwann cells (Toma et al 2007;Santos-Silva and Cavalcante 2001;Ramón-Cueto and Nieto-Sampedro 1992).…”

“…The similar regulation pattern of CNPase expression in both cell types is a further evidence for a similar functional role in both cell types. Since it is known that OPCs in vitro express CNPase under differentiating but not under proliferative conditions (Pringproa et al 2008), it is conceivable to speculate that CNPase in OECs and Schwann cells is related to cellular differentiation.…”