Non-viral gene delivery transfection profiles influence neuronal architecture in an in vitro co-culture model.
Non-viral gene delivery transfection profiles influence neuronal architecture in an in vitro co-culture model.
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DOI：
10.1002/bit.22311
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发表时间：
2009-08-01
影响因子：
3.8
通讯作者：
Shea, Lonnie D.
中科院分区：
文献类型：
--
作者：
Houchin-Ray, Tiffany;Zelivyanskaya, Marina;Huang, Alyssa;Shea, Lonnie D.
Gene delivery from tissue engineering scaffolds can induce expression of tissue inductive factors to stimulate the cellular processes required for regeneration. Transfected cells secrete diffusible proteins that can create local concentration gradients, depending on the number, distribution, and expression level of transfected cells. These gradients are linked to cellular organization and tissue architecture during embryogenesis. In this report, we investigate neuronal architecture and neurite guidance in response to the concentration gradients achieved by localized secretion of a neurotrophic factor from transfected cells. A co-culture model was employed to examine neuronal responses to multiple transfection profiles, which affects the local concentration of secreted nerve growth factor (NGF). Neuronal architecture, as defined by number of neurites per neuron and length of neurites, was influenced by the transfection profile. Low levels of NGF production by few transfected cells produced longer primary neurites with less branching relative to the higher expression levels or increased numbers of transfected cells. Furthermore, for low NGF production by few transfected cells, the growth cone of the axons was marked by longer extensions and larger surface area, suggesting the presence of a guidance cue. Control studies with varying NGF concentrations did not substantially alter the neuronal architecture, further supporting an effect of localized concentration gradients, and not simply the concentration. Mathematical modeling of NGF diffusion was employed to predict the concentration gradients produced by the transfection profiles, and the resultant gradients were correlated to the cellular response. This report connects the transfection profile, concentration gradients, and the resulting cellular architecture, suggesting a design consideration for the application of gene delivery to regenerative medicine.
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