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Application of aurintricarboxylic acid for the adherence of mouse P19 neurons and primary hippocampal neurons to noncoated surface in serum‐free culture
Authors:Tsung‐Yih Lee  Wen‐Shin Chen  Yung‐An Huang  Ting‐Wei Liu  Eric Hwang  Ching‐Ping Tseng
Affiliation:1. Dept. of Biological Science and Technology, National Chiao Tung University, Hsinchu, 30068, Taiwan;2. Institute of Bioinformatics and Systems Biology, National Chiao Tung University, Hsinchu, 30068, Taiwan;3. Institute of Molecular Medicine and Bioengineering, National Chiao Tung University, Hsinchu, 30068, Taiwan
Abstract:
Dissociated primary neuron culture has been the most widely used model systems for neuroscience research. Most of these primary neurons are cultured on adhesion matrix‐coated surface to provide a proper environment for cell anchorage under serum‐free conditions. In this study, we provide an alternative technique to promote the adhesions of these neurons using aurintricarboxylic acid (ATA), a nonpeptide compound, without surface manipulations. We first demonstrated that ATA could promote Chinese hamster ovary cell attachment and proliferation in serum‐free medium in a dosage‐dependent manner. We later showed that ATA significantly enhanced the attachment of the retinoic acid differentiated P19 mouse embryonal carcinoma (P19) neurons, with an optimal concentration around 30 μg/mL. A similar result was seen in primary hippocampal neurons, with an optimal ATA concentration around 15 μg/mL. Further morphological assessments revealed that the average neurite length and neuronal polarization were almost identical to that obtained using a conventional method with poly‐L ‐lysine surface. The advantages of using the ATA treatment technique for immunochemical analysis are discussed. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012
Keywords:aurintricarboxylic acid  serum‐free medium  P19 neuron cells  hippocampal neurons  cell attachment reagent
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