Uncorrected Proof Arch Neurosci. In Press(In Press):e99771. Published online 2020 March 29. doi: 10.5812/ans.99771. Brief Report Primary Spinal Cord Neurons Better Attach to Non-Coated Surfaces Than to Poly(D-lysine)-Coated Surfaces Fateme Azizi 1 , Rasoul Ghasemi 2 and Mahmoudreza Hadjighassem 3, 4, * 1 School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran, Iran 2 Department of Physiology and Neurophysiology Research Center, Faculty of Medicine, Shahid Beheshti Medical Sciences University, Tehran, Iran 3 Brain and Spinal Cord Injury Research Center, Neuroscience Institute, Tehran University of Medical Sciences, Tehran, Iran 4 Department of Neuroscience and Addiction Studies, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran, Iran * Corresponding author: MD, Ph.D., Brain and Spinal Cord Injury Research Center, Neuroscience Institute, Tehran University of Medical Sciences, Tehran, Iran. Email: mahmoudreza@hotmail.com Received 2019 November 28; Revised 2020 January 16; Accepted 2020 February 25. Abstract Background: Cell culture is an important technique in cellular and molecular biology. There are two basic systems for culturing cells, adherent (monolayer) culture and suspension culture. In adherent cell culture, cellular adhesion molecules play a vital role in many physiological processes. Many cells, especially primary-obtained cells, hardly attach to plates, so coating agents are used for cell attachment and growth. There is no striking report on whether all cells need coated plates. Objectives: We investigated whether a coating plate is essential for primary spinal cord-obtained neuron culture. Methods: Cells were separated and seeded in tissue culture plates coated with either Poly(L-lysine) or Poly(D-lysine), as well as with- out coating containing DMEM-F12 media with 10% FBS. Results: Interestingly, we found that neuron cells more potently attached to coating free plates. Conclusions: These results provide strong evidence helping the researchers to optimize primary spinal cord cultures. Keywords: Neurons, Primary Cell Culture, Poly(L-lysine), Spinal Cord, Cell Adhesion 1. Background Cell culture has become one of the most fundamen- tal techniques for modeling biological systems (1). Gener- ally, cell culture is categorized into adherent (monolayer) culture and suspension culture. In adherent cell culture, which is the abundant form of culture, it provides the an- chorage of cells to growth surfaces (2). However, the ad- hesion properties of the cells are cell-dependent; while some cells could attach per se, some others need adhe- sion molecules to assist their anchorages. Conventionally, it is believed that neuronal cells need adhesion molecules to promote their attachment; therefore, coated plates are used for this purpose (3). However, it is not clearly specified if all types of neurons need such assistance. In this short report, we compare primary spinal cord cultures in two different situations: non-coated polystyrene plates and plates coated with agents (Poly(D- lysine) and Poly(L-lysine)). 2. Objectives We investigated whether the coating is essential for culturing of primary spinal cord-obtained neurons. 3. Methods A neonatal rat was anesthetized with ether. The spinal cord was extruded from the neonatal rat using hydraulic pressure. The obtained spinal cord was transferred to a tube containing ice-cold PBS and antimycotic antibiotic, then dissociated into a cell suspension by mechanical dis- ruption and trypsinization (0.125% trypsin). Cells were sep- arated and seeded at a density of 1 × 10 6 cells/mL in tissue culture plates coated with either Poly(L-lysine) or Poly(D- lysine), as well as without coating containing DMEM-F12 media with 10% FBS. 3.1. Coating Plates were coated with each agent for 2 hours at 37°C, then washed 2 - 3 times with PBS to minimize the toxic ef- fect of the coating material. 4. Results We found that obtained cells better attached to polystyrene plates (without coating) and grew well. After three days of in vitro culture, the number of adhered Copyright © 2020, Author(s). This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.