Apr 19, 2026
Polyamine Pathway Modulation in 2D Monoculture
- 1Icahn School of Medicine at Mount Sinai
Protocol Citation: Elena Coccia 2026. Polyamine Pathway Modulation in 2D Monoculture. protocols.io https://dx.doi.org/10.17504/protocols.io.kqdg3mo8ql25/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: April 19, 2026
Last Modified: April 19, 2026
Protocol Integer ID: 315320
Keywords: polyamine, spermidine, AMXT-1501, cell growth, protein expression, 2D monoculture, polyamine pathway modulation in 2d monoculture, polyamine pathway modulation, effects of polyamine pathway modulation, protein expression in 2d monoculture cell system, 2d monoculture cell system, using spermidine, 2d monoculture
Disclaimer
This protocol was generated by AI
Abstract
To investigate the effects of polyamine pathway modulation using spermidine and AMXT-1501 on cell growth and protein expression in 2D monoculture cell systems.
Materials
Cell Culture Plates (6-well plates) - 6 plates
Growth Media (DMEM supplemented with 10% FBS and antibiotics) - 30 mL
Sterile Water (For preparing spermidine stock) - 10 mL
DMSO (For preparing AMXT-1501 stock) - 10 mL
Reagents:
Polyamine Spermidine - A polyamine involved in cellular growth and function
Inhibitor AMXT-1501 - A compound used to modulate polyamine metabolism
Troubleshooting
Problem
Cells do not adhere to plates
Solution
Ensure plates are properly coated and cells are not over-trypsinized.
Problem
Low cell viability post-treatment
Solution
Check concentrations of spermidine and AMXT-1501; ensure proper media refreshment.
Safety warnings
Handle all reagents and materials according to safety data sheets. Use personal protective equipment (PPE) including gloves and lab coats. Work with DMSO and spermidine in a fume hood to avoid inhalation.
Cell Culture Preparation
Culture cells in standard 2D monoculture conditions using appropriate growth media.
Treatment Application
Prepare spermidine and AMXT-1501 treatments. For spermidine: Prepare stock in water at a 10mM stock in DMEM. For AMXT-1501: Prepare stock in DMSO at 10mM.
Treatment Addition
Replace existing media in wells with treatment media (spermidine or AMXT-1501). Control wells should receive equivalent volumes of vehicle (water for spermidine; DMSO for AMXT-1501).
Media Refreshment
Refresh media containing treatments every other day for the duration of the experiment. Ensure to maintain the same treatment concentrations.
Protocol references