Enrichment of rat oligodendrocyte progenitor cells by magnetic cell sorting

RETRACTED ARTICLE: Pericyte degeneration causes white matter dysfunction in the mouse central nervous system

“…A2B5-positive oligodendrocyte precursor cells (OPCs) were isolated from cortices of 129S1/Svlmj P3-P6 mouse pups by magnetic cell sorting (MACS, Miltenyi Biotec), as previously described 73 . Briefly, brains were removed, minced, and further processed using Neural Dissociation Kit following the manufacturer's instructions (#130-092-628, Miltenyi Biotec).…”

“…OPCs were plated at a density of 5×10 4 cells on poly-D-lysine (PDL)-coated coverslips for cell viability and immunocytochemistry experiments or on PDL-coated 6-well dishes for western blotting and caspase-3 activity assays. OPCs were allowed to differentiate into mature oligodendrocytes in medium containing DMEM-SATO based growth medium+Forskolin+NT3+T3 (T3 is the active hormone, 3,5,30-tri-iodothyronine), which has been shown to promote oligodendrocyte differentiation from OPCs 73,74 . Experiments were performed on 7 days in vitro (DIV) mature oligodendrocyte cultures that were approximately 90% positive for myelin basic protein (MBP, a marker of mature oligodendrocytes) 74 and negative for O4, a marker for an intermediate transitional cell type (pre-immature oligodendrocytes) between OPCs and mature oligodendrocytes.…”

Stem Cell Glycolipids

“…A method to isolate O-2A progenitor cells in nerve tissues by immunopanning using antibodies to rat neural antigen-2, galactosylceramide (for negative selection), and A2B5 antigens (for positive selection) has been widely used [101]. It has also been reported that it is possible to enrich rat oligodendrocyte progenitor cells in a heterogeneous population of neural stem cells by magnetic cell sorting using A2B5 antibody [102].…”

Mesenchymal Stromal Cells and Neural Stem Cells Potential for Neural Repair in Spinal Cord Injury and Human Neurodegenerative Disorders

“…These contradictory findings are often explained in regard to beneficial (in vitro) or inhibitory (in vivo) conditions directly influencing neuronal or glial fate (Weiss et al 1996). Furthermore, after pathological condition such as spinal cord injury, most of the newly dividing intrinsic ependymal stem cells migrate toward damaged tissue, where they develop into macroglial cells, while only few cells retain primitive nestin-like phenotype (Johansson et al 1999;Cizkova et al 2009a). Likewise, a significant number of neural progenitors could be activated also in other regions of the parenchyma ) (Kehl et al 1997).…”

“…In particular, magnetic cell separation using specific monoclonal antibodies (e.g. A2B5, PSA-NCAM) conjugated to nanoparticles allowing positive retention or negative dilution of selected cells provide a feasible approach for experimental cell enrichment of desired oligodendroglial progeny, which may be used in future trials for cell-based therapies to treat spinal cord injury (Cizkova et al 2009a). These studies have shown that MACs technology enable us to gain about a 5 to 9 fold increase of immature, mature oligodendrocytes content (NG2+, RIP+, MBP+) when compared to amount of oligodendroglial cells acquired from unseparated population (Fig.2).…”