title: Application of aurintricarboxylic acid for the adherence of mouse P19 neurons and primary hippocampal neurons to noncoated surface in serum-free culture.
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.
authors.affiliation: Dept. of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan.
authors.forename: Tsung-Yih
authors.initials: TY
authors.lastname: Lee
authors.forename: Wen-Shin
authors.initials: WS
authors.lastname: Chen
authors.forename: Yung-An
authors.initials: YA
authors.lastname: Huang
authors.forename: Ting-Wei
authors.initials: TW
authors.lastname: Liu
authors.forename: Eric
authors.initials: E
authors.lastname: Hwang
authors.forename: Ching-Ping
authors.initials: CP
authors.lastname: Tseng
mesh_terms: D000199:Actins / Q000378:metabolism; D000704:Analysis of Variance; D000818:Animals; D001312:Aurintricarboxylic Acid / Q000494:pharmacology*; D016466:CHO Cells; D002448:Cell Adhesion / Q000187:drug effects*; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D048430:Cell Shape / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D016895:Culture Media, Serum-Free; D006624:Hippocampus / Q000166:cytology* / Q000187:drug effects*; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000166:cytology* / Q000187:drug effects*; D013499:Surface Properties; D014212:Tretinoin / Q000494:pharmacology; D014404:Tubulin / Q000378:metabolism
chemical_list: D000199:Actins; D016895:Culture Media, Serum-Free; D014404:Tubulin; D001312:Aurintricarboxylic Acid; D014212:Tretinoin