Differentiation of a human neuroblastoma into neuron-like cells increases their susceptibility to transduction by herpesviral vectors.
Gene transfer is a powerful tool for functional gene analysis in human cells. In this respect, there is a need to develop experimental models that involve homogeneous cultures of human neuron-like cells susceptible to gene transduction and that are easy to handle. Here we describe an optimized and reproducible procedure to differentiate human SH-SY5Y neuroblastoma cells into a homogeneous population of neuron-like cells. The fully differentiated cells are postmitotic and resemble primary cultured neurons in terms of their cytoskeletal polarity. Notably, differentiated SH-SY5Y cells are far more susceptible to transduction by herpes simplex virus (HSV-1)-based vectors than proliferating SH-SY5Y cells. This increase in transduction efficiency after neuronal differentiation may be due to the up-regulation of cell surface receptors for herpesvirus entry. In summary, we propose that fully differentiated human neuron-like cells obtained from the SH-SY5Y neuroblastoma may constitute an excellent and versatile experimental tool for gene transfer and functional genomic studies with HSV-1 vectors.
Gimenez-Cassina
Alfredo
A
Departamento de Biologia Molecular, Centro de Biologia Molecular Severo Ochoa, Universidad Autonoma de Madrid, Madrid, Spain.
Lim
Filip
F
Diaz-Nido
Javier
J
D015153:Blotting, Western / Q000379:methods; D001973:Bromodeoxyuridine; D002454:Cell Differentiation / Q000502:physiology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D004198:Disease Susceptibility / Q000821:virology; D005455:Fluorescent Antibody Technique / Q000379:methods; D015870:Gene Expression / Q000502:physiology; D005822:Genetic Vectors / Q000502:physiology*; D006801:Humans; D007211:Indoles; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009447:Neuroblastoma; D016900:Neurofilament Proteins / Q000378:metabolism; D009474:Neurons / Q000502:physiology* / Q000821:virology*; D018139:Simplexvirus / Q000502:physiology*; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods*
D007211:Indoles; C489788:MAP2 protein, human; D008869:Microtubule-Associated Proteins; D016900:Neurofilament Proteins; C007293:DAPI; D001973:Bromodeoxyuridine