Anti-A2B5 Microbeads from Miltenyi Biotec

The mammalian nervous system is comprised of several types of cells including neurons and glial cells. Neurons perform the primary function of sending and receiving signals while glial cells are support cells. They perform several important functions in the nervous system, including providing structural and nutritional support to the neurons. There are several types of glial cells including astrocytes, oligodendrocytes, microglia and Schwann cells. Neural stem cells are defined as cells that are capable of differentiating into neurons as well as glia. Committed precursor or progenitor cells are those whose lineage is committed to either a neuronal or glial lineage. A2B5 is a marker that is used to identify glial-committed progenitor cells. A2B5 is a C-series ganglioside antigen expressed by glial restricted progenitors. It is expressed predominantly in embryonic and neonatal tissue. These cells are shown to be capable of differentiating in to both oligodendrocytes and astrocytes. Several studies have used expression of A2B5 to isolate immature glial committed progenitor cells.

Miltenyi Biotec have developed Anti-A2B5 MicroBeads for positive isolation of A2B5 expressing cells from a variety of tissues and cell populations, including mouse SVZ tissue and fetal human brain tissue. The kit contains magnetic microbeads labeled with anti-A2B5 antibodies and an FcR blocking agent to block non-specific binding. The protocol for isolation is simple and is not time consuming. The cells are first magnetically labeled with A2B5 microbeads by incubating the cells for 10 min with the blocking agent and for 15 min with the microbeads at 4°C. The A2B5+ cells are subsequently separated by passing the cell suspension through a column placed in the magnetic field of a MACS cell separator. Once the unlabelled A2B5 negative cells flow through, the column is removed from the magnetic field and the A2B5+ cells are flushed out.

The A2B5 MicroBead Kit has been used in our laboratory for the past year for isolation of glial-committed progenitor cells from fetal human brains on several occasions. The procedure for this kit is relatively easy and a reasonably pure population of A2B5 expressing cells can be isolated from a heterogeneous cell suspension. Since the procedure is not too long and since most reactions are done on ice, cell viability is not compromised. As with other kits from Miltenyi Biotec, to use this kit effectively, it is necessary to purchase magnetic cell separators: MiniMACS, MidiMACS or SuperMACS, and positive selection columns from the company. The company also offers a PE conjugated A2B5antibody for flow cytometric or fluorescence microscopic verification of the isolated cells.

In summary, the Anti-A2B5 MicroBeads from Miltenyi Biotec are a very convenient tool for isolation of A2B5 expressing progenitor cells from rodent and human tissues.