The human neuroblastoma cell line, SH-SY5Y cells, derived from the parental SK-N-SH cell line, is commonly used as an in vitro model for neuroscience and neurobiology research. Since SH-SY5Y cells are widely cultured for research, several different culture media have been used to optimize the growth of the cells, including Eagle's Minimum Essential Medium (EMEM), Dulbecco’s modified Eagle’s medium (DMEM) and other recently developed culture media. SH-SY5Y cells has the ability to reach confluency in culture flasks ranges from 5 days to 15 days, depending on the culture media used. Hence, the optimization of the culture media is crucial to achieve the fastest growth rate for the cells. The objective of the study is to evaluate the culture media for the proliferation of SH-SY5Y cells. We compared the growth rate of SH-SY5Y cells cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 15% heat-inactivated fetal bovine serum (hiFBS), Dulbecco’s modified Eagle’s medium: Nutrient mixture F-12 (DMEM:F12) + supplemented with 15% hiFBS and DMEM:F12 supplemented with 10% hiFBS. In DMEM:F12 supplemented with 15% hiFBS, cells grew up to 6.67E+05 cells. In DMEM:F12 supplemented with 10% hiFBS, cells grew up to 5.28E+05 cells. In DMEM supplemented with 15% hiFBS, the cells grew up to 4.76E+05 cells. There was a significant difference between culture media DMEM:F12 supplemented with 15% hiFBS as compared to DMEM:F12 supplemented with 10%hiFBS and DMEM supplemented with 15% hiFBS (p0.05). We found that DMEM:F12 supplemented with 15% hiFBS could serve as an optimized culture media for high proliferation rate of SH-SY5Y cells.
Nur Khalisah Kaswan, Nurul Syazwani Mohd Suhaimi, Noor Aishah Mohammed Izham, Tengku Azam Shah Tengku Mohamad, Mohd Roslan Sulaiman, Enoch Kumar Perimal
Background: Cardamonin is a naturally occurring chalcone from the Alpinia species. It is known to possess antioxidant and anti-inflammatory properties. Our previous studies have shown that cardamonin has antihyperalgesic and antiallodynic effects on CCI-induced neuropathic pain in mice. Although the evidence of the association between cardamonin and neuropathic pain has been reported in animal studies, specific targets using in vitro models are still lacking. Objectives/Methods: This study aims to investigate the effect of cardamonin on nitric oxide production using the LPS-induced neuropathic pain-like SH-SY5Y in vitro model through NMDA receptor expression. Results: Cardamonin administration in differentiated SH-SY5Y cells significantly reduced nitric oxide production assessed using Griess reagent. Western blot analysis demonstrated a significant reduction in GluN2B receptor expression in the cardamonin treated SHSY5Y cells compared to the vehicle treated group. Conclusions: These data suggest that cardamonin reduces nitric oxide production modulated through NMDA GluN2B receptor subunit. Our results provides preliminary data to support the in vivo studies using cardamonin and may contribute to further understanding the mechanisms of action of cardamonin.