Development of an artificial neuronal network with post-mitotic rat fetal hippocampal cells by polyethylenimine.
The selection of appropriate surface materials that promote cellular adhesion and growth is an important consideration when designing a simplified neuronal network in vitro. In the past, extracellular matrix proteins such as laminin (LN) or positively charged substances such as poly-l-lysine (PLL) have been used. In this study, we examined the ability of another positively charged polymer, polyethyleneimine (PEI), to promote neuronal adhesion, growth and the formation of a functional neuronal network in vitro. PEI, PLL and LN were used to produce grid-shape patterns on glass coverslips by micro-contact printing. Post-mitotic neurons from the rat fetal hippocampus were cultured on the different polymers and the viability and morphology of these neurons under serum-free culture conditions were observed using fluorescent microscopy and atomic force microscopy (AFM). We show that neurons cultured on the PEI- and PLL-coated surfaces adhered to and extended neurites along the grid-shape patterns, whereas neurons cultured on the LN-coated coverslips clustered into clumps of cells. In addition, we found that the neurons on the PEI and PLL-coated grids survived for more than 2 weeks in serum-free conditions, whereas most neurons cultured on the LN-coated grids died after 1 week. Using AFM, we observed some neurosynapse-like structures near the neuronal soma on PEI-coated coverslips. These findings indicate that PEI is a suitable surface for establishing a functional neuronal network in vitro.
Liu
Bingfang
B
Beijing Institute for Neuroscience, Beijing Center for Neural Regeneration and Repairing, Key Laboratory for Neurodegenerative Diseases of the Ministry of Education, Capital Medical University, Beijing 100069, PR China.
Ma
Jun
J
Gao
Erjing
E
He
Yu
Y
Cui
Fuzhai
F
Xu
Qunyuan
Q
D000200:Action Potentials / Q000502:physiology*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D006624:Hippocampus / Q000166:cytology* / Q000196:embryology* / Q000502:physiology; D009415:Nerve Net / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D011094:Polyethyleneimine / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley
D011094:Polyethyleneimine