Purification of embryonic stem cell-derived neurons by immunoisolation.
The pluripotency and high proliferative capacity of embryonic stem (ES) cells (1-3) makes them an attractive source of different cell types for biomedical research and cell replacement therapies. A major prerequisite for these applications is the availability of a homogeneous population of the desired cell type. However, ES cell-derived material contains, for example, undifferentiated cells, which can cause tumor formation after transplantation into the brain (4). To avoid such unwanted side effects, effective purification of distinct types of cells needs to be developed. Here, we describe an immunoisolation procedure to purify neurons from in vitro differentiated mouse ES cells using an antibody against the neuronal cell adhesion molecule L1 (5, 6). Our procedure yields a pure population of differentiated neurons, which are electrically excitable and form excitatory, glutamatergic, and inhibitory GABAergic synapses. The ability to highly purify ES cell-derived neurons will boost their molecular characterization and the further exploration of their therapeutic potential.
D000200:Action Potentials; D000818:Animals; D000911:Antibodies, Monoclonal; D002454:Cell Differentiation; D004622:Embryo, Mammalian / Q000166:cytology*; D007150:Immunohistochemistry; D051379:Mice; D039842:Neural Cell Adhesion Molecule L1 / Q000276:immunology; D016900:Neurofilament Proteins / Q000276:immunology / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D018408:Patch-Clamp Techniques; D011963:Receptors, GABA-A / Q000502:physiology; D017470:Receptors, Glutamate / Q000502:physiology; D013234:Stem Cells / Q000502:physiology*; D013569:Synapses / Q000502:physiology
D000911:Antibodies, Monoclonal; D039842:Neural Cell Adhesion Molecule L1; D016900:Neurofilament Proteins; D011963:Receptors, GABA-A; D017470:Receptors, Glutamate; C051774:neurofilament protein H