PNS Cell Protocols
Change Log
Protocol | Version/ No. | Section | Change log | Rationale | Previous version | New version |
Sensory Neuron Progenitors | COM/PRT/31 Dec 2023 / version 1 | Preparation of Media and Coating Reagents | SureBond-XF addition to Neural Plating Media | To aid cell adherence | No SureBond-XF addition | |
Sensory Neuron Progenitors | COM/PRT/31 Dec 2023 / version 1 | Preparation of Media and Coating Reagents | Mitomycin C concentration and reconstitution | Optimised dosing to prevent neuronal cell death | 2.5µg/ml
Made up in DMSO* |
2.0µg/ml
Made up in water (so needs to be made fresh each time) |
Sensory Neuron Progenitors | COM/PRT/31 Dec 2023 / version 1 | Preparation of Media and Coating Reagents | Lyophilized PDL changed to liquid PDL and change in concentration used | Enables improved cell adherence and plating consistency | Powdered PDL made up in water at and used at 0.1mg/ml | Liquid PDL diluted to 0.05mg/ml |
Sensory Neuron Progenitors | COM/PRT/31 Dec 2023 / version 1 | Coating Cell Culture Ware | Coating incubation time and temperature | Enables improved cell adherence and plating consistency | Incubate plate coated with PDL for 5 mins at room temperature | Incubate plate coated with PDL for 60 mins at 37°C |
Sensory Neuron Progenitors | COM/PRT/31 Dec 2023 / version 1 | Differentiation of axoCells Sensory Neuron Progenitors | Seeding density | Optimised cell density based on new coating reagents | 100,000/cm2 | 100,000 – 150,000/cm2 |
* NB. You can revert to the previous method and make up mitomycin C in DMSO (which can be stored for up to 4 weeks at -80°C) but performance in Axol’s hands is better when made up in water.
Protocol | Version/ No. | Section | Change log | Rationale | Previous version | New version |
Motor Neuron Progenitors | COM/PRT/20 Dec 2023 / version 1 | Preparation of Media and Coating Reagents | Lyophilized PDL changed to liquid PDL and change in concentration used | Enables improved cell adherence and plating consistency | Powdered PDL made up in water and used at 0.1mg/ml | Liquid PDL diluted to 0.05mg/ml |
Motor Neuron Progenitors | COM/PRT/20 Dec 2023 / version 1 | Coating Cell Culture Ware | Coating incubation time and temperature | Enables improved cell adherence and plating consistency | Incubate plate coated with PDL for 5 mins at room temperature | Incubate plate coated with PDL for 60 mins at 37°C |
Motor Neuron Progenitors | COM/PRT/20 Dec 2023 / version 1 | Differentiation of axoCells Motor Neuron Progenitors | Seeding density | Optimised cell density based on new coating reagents | 150,000/cm2 | 130,000 – 180,000/cm2 |