User Guide

Quick-NeuronTM Cholinergic - Maintenance Medium
Catalog Number: CH-MMIntroduction
Quick-Neuron™ Cholinergic - Maintenance Medium may be used for the long-term maintenance of human pluripotent
stem cell-derived cholinergic neurons following differentiation as outlined in the Quick-Neuron™ Cholinergic - SeV Kit,
mRNA Kit, and Human iPSC-derived Neurons user guides. Quick-Neuron™ Cholinergic differentiated cell cultures display typical neurite outgrowth and express a variety of neuronal markers, such as the pan-neuronal marker tubulin beta
3 class III (TUBB3) and the cholinergic neuron marker choline acetyltransferase (ChAT). When handled and maintained
according to the instructions in this user guide, cholinergic neurons are viable long-term and are suitable for a variety ofcharacterization and neurotoxicity assays.
Scale: 					The Quick-Neuron™ Cholinergic - Maintenance Medium provides sufficient medium for 4		
wells of a 24-well plate for up to 2 weeks.

Related Products:						Quick-Neuron™ Cholinergic - SeV Kit, Catalog Number: CH-SeV
Quick-Neuron™ Cholinergic - mRNA Kit, Catalog Number: CH-mRNA
Quick-Neuron™ Cholinergic - Human iPSC-derived Neurons, Catalog Number: CH-SeV-CW

Kit Contents
Upon receipt, store the reagents at the temperatures indicated in the table below. All reagents are shipped on dry ice.
Reagents

VolumeStorage

Component N1

830 μl

-20°C or -80°C

Component B

16 μl-20°C or -80°C

Component P

14 μl-20°C or -80°C

Required Consumables
Item

VendorCatalog Number

DMEM/F12

ThermoFisher

21331020

Neurobasal Medium

ThermoFisher

21103049

Glutamax (100x)

ThermoFisher

35050061

Penicillin-Streptomycin

ThermoFisher

15140122

Conditions of Use
This product is for research use only. It is not approved for use in humans or for therapeutic or diagnostic use.

Technical Support
For technical support, please contact us at cs@elixirgensci.com or call +1 (443) 869-5420 (M-F 9 am-5 pm EST).
Last revised: January 22, 2021

elixirgensci.com

1Base Media Preparation
Medium N1
1. Prepare Medium N1 using the reagents listed in the table below.
• Thaw Component N1 at 4°C overnight or 30 minutes on ice.
• All other reagents should be warmed at room temperature for 20-30 minutes.
Medium N1 Reagents

VolumeDMEM/F128 ml

Neurobasal Medium8 ml

200 mM Glutamax (100x)

83 μl

Penicillin-Streptomycin (10000 units/ml; 100x)

167 μlComponent N1

500 μl

2. Store Medium N1 for up to 2 weeks at 4°C.
• The leftover Component N1 can be discarded or saved for another use.

First Week
Medium N1(BP)1. Prepare Medium N1(BP) using the reagents listed in the table below.
• Thaw Component B at 4°C overnight or 30 minutes on ice. Spin down before use.
• All other reagents should be warmed at room temperature for 20-30 minutes.
Medium N1(BP) Reagents

VolumeMedium N1

7 ml

Component B

7 μl

Component P

3.5 μl2. Save the leftover Component B at 4°C.
• The leftover Component P can be discarded or saved for another use.
3. Warm Medium N1(BP) at room temperature for 20-30 minutes until it no longer feels cold.
4. Pipet out most of the old medium, but not completely (i.e., just enough to cover the surface of the well), from eachwell using a P1000 pipettor and add 800 μl Medium N1(B) along the wall of the well very slowly.
5. Incubate the cultures at 37°C, 5% CO2 for 2 days.
6. For subsequent medium changes, pipet out half (400 μl) of the old medium from each well using a P1000 pipettor
and add 400 μl Medium N1(BP).
7. Repeat Step 6 every 2-3 days such as on Monday, Wednesday, and Friday for 1 week.

2elixirgensci.com

Second Week
Medium N1(B)
1. Prepare fresh Medium N1(B) using the reagents listed in the table below.
• Warm Medium N1 at room temperature for 30 minutes.
• Place Component B on ice. Spin down before use.
Medium N1(B) Reagents

VolumeMedium N1

7 ml

Component B

7 μl

2. Warm Medium N1(B) at room temperature for 20-30 minutes until it no longer feels cold.
3. Pipet out most of the old medium, but not completely (i.e., just enough to cover the surface of the well), from eachwell using a P1000 pipettor and add 800 μl Medium N1(B) along the wall of the well very slowly.
4. Incubate the cultures at 37°C, 5% CO2 for 2 days.
5. For subsequent medium changes, pipet out half (400 μl) of the old medium from each well using a P1000 pipettor
and add 400 μl Medium N1(B).
6. Repeat Step 5 every 2-3 days such as on Monday, Wednesday, and Friday for 1 week.

elixirgensci.com

3