PNS Cell Protocols

View and download Axol’s validated user protocols for iPSC-derived PNS cells

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