Abstract
Myelination of neuronal axons is essential for proper brain functioning and requires mature myelinating oligodendrocytes (myOLs). The human OL cell lines HOG and MO3.13 have been widely used as in vitro models to study OL (dys) functioning. Here we applied a number of protocols aimed at differentiating HOG and MO3.13 cells into myOLs. However, none of the differentiation protocols led to increased expression of terminal OL differentiation or myelin-sheath formation markers. Surprisingly, the applied protocols did cause changes in the expression of markers for early OLs, neurons, astrocytes and Schwann cells. Furthermore, we noticed that mRNA expression levels in HOG and MO3.13 cells may be affected by the density of the cultured cells. Finally, HOG and MO3.13 co-cultured with human neuronal SH-SY5Y cells did not show myelin formation under several pro-OL-differentiation and pro-myelinating conditions. Together, our results illustrate the difficulty of inducing maturation of HOG and MO3.13 cells into myOLs, implying that these oligodendrocytic cell lines may not represent an appropriate model to study the (dys)functioning of human (my)OLs and OL-linked disease mechanisms.
Highlights
In the central nervous system, axons are ensheathed by myelin which supports neuronal conduction velocity, and provides metabolic and trophic support to neurons [1]
In none of the conditions, mRNA expression of the early OL precursor cells (OPCs) marker VCAN was found (Figure 1A–C), while the astrocyte marker FGFR3 was expressed in undifferentiated HOG cells and still expressed after differentiation (Figure 1A–C)
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Summary
In the central nervous system, axons are ensheathed by myelin which supports neuronal conduction velocity, and provides metabolic and trophic support to neurons [1]. OLs (imOLs) and mature OLs (mOLs) into myelinating OLs (myOLs) [1,2,3]. The ‘early-myelination stage’ is specified by the expression of myelin basic protein (MBP), proteolipid protein 1 (PLP1), 20 ,30 -Cyclic-nucleotide 30 -phosphodiesterase (CNP) and UDP-galactose ceramide (UGT8A), and the ‘late stage’ by myelin-associated glycoprotein (MAG), myelin-oligodendrocyte basic protein (MOBP), myelin-oligodendrocyte glycoprotein (MOG), myelin and lymphocyte protein (MAL), aspartoacylase (ASPA) and plasmolipin (Tm4sf11) expression [4]. The ‘early-myelination stage’ genes will be referred here to as mOL markers and the ‘late-stage’ genes as myOL markers. For in vitro functional studies on OLs, the human oligodendrocytic cell lines HOG and MO3.13 are often used. The HOG cell line is directly derived from an oligodendroglioma [5] and the MO3.13
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