User Guide
Quick-Neuron™ Cholinergic - Maintenance Medium
Catalog Number: CH-MMIntroduction
Quick-NeuronTM 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-NeuronTM Cholinergic - SeV Kit and HumaniPSC-derived Neurons user guides. Quick-NeuronTM Cholinergic differentiated cell cultures display typical neurite outgrowthand express a variety of neuronal markers, such as the pan-neuronal marker tubulin beta 3 class III (TUBB3) and thecholinergic marker choline acetyl-transferase (ChAT). When handled and maintained according to the instructions in thisuser guide, cholinergic neurons are viable long-term and are suitable for a variety of characterization and neurotoxicity
assays.
Scale:The Quick-Neuron™ Cholinergic - Maintenance Medium provides sufficient medium for 4 wells ofa 24-well plate, 1 well of a 6-well plate, or 16 wells of a 96-well plate for up to 2 weeks.

Related Products:

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

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

VolumeStorage

Thaw

Component N1

830 μl

-20°C or -80°C

On ice or 4°C

Component B

20 μl-20°C or -80°C

On ice or 4°C

Component P

50 μl-20°C or -80°C

Room temperature

Condition 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 refer to the FAQ on our website.
You may also contact us at cs@elixirgensci.com or call +1 (443) 869-5420 (M-F 9am-5pm EST).

Last revised: July 14, 2023

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1Required Consumables
Item

VendorCatalog Number

DMEM/F12

ThermoFisher

21331020

Neurobasal Medium

ThermoFisher

21103049

GlutaMAX

ThermoFisher

35050061

Penicillin-Streptomycin

ThermoFisher

15140122

Preparation
Medium N1(B)1.

Prepare Medium N1(B) using the reagents listed in the table below.
○ Thaw Components N1 and B for 20-30 minutes at the temperature indicated in the “Contents” table on
page 1.
○ Warm all other reagents at room temperature for 20-30 minutes.
○ Tap each Component tube 3 times and then briefly spin all tubes down before use.
○ Keep Medium N1(B), and any subsequent media made with it, protected from light.
○ Store Medium N1(B) for up to 2 weeks at 4°C.
○ Leftover Component N1 can be discarded or saved at 4°C for up to two weeks.
Reagents

VolumeDMEM/F12

7.2 mlNeurobasal Medium

7.2 mlGlutaMAX

75 μl

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

150 μlComponent N1

450 μlComponent B

15 μl

First Week
1.

Prepare Medium N1(BP) using the reagents listed in the table below.
○ Thaw Component P for 20-30 minutes at the temperature indicated in the “Contents” table on page 1.
○ Warm all other reagents at room temperature for 20-30 minutes.
○ Tap the Component P tube 3 times and then briefly spin it down before use.
○ Store Medium N1(BP) for up to 2 weeks at 4°C.
○ Leftover Component P can be saved at 4°C.
Reagents

VolumeMedium N1(B)

7 ml

Component P

3.5 μlelixirgensci.com

22. Pipet out most of the old medium, but not completely (i.e., just enough to cover the surface of the well), from eachwell and very slowly along the wall of the well, add room temperature Medium N1(BP) according to the following
table.
Required volume per well
Reagents
Medium N1(BP)6-well plate24-well plate

96-well plate

2 ml

800 µl150 µl3. Incubate the cultures at 37°C, 5% CO2 for 2 days.
4. For subsequent medium changes, pipet out half (see volumes in the table above) of the old medium from each wellusing and replace with an equal volume of room temperature Medium N1(BP).
5. Repeat Step 4 every 2-3 days such as on Monday, Wednesday, and Friday for 1 week.

Second Week
1.

Warm Medium N1(B) at room temperature for 20-30 minutes until it no longer feels cold.
Note: If there is an outgrowth of non-neuronal flat cells in the culture (as seen marked by arrows in the sample
image below) users should continue using Medium N1(BP) in the second week, following the instructions toprepare Medium N1(BP) provided in the “First Week” .

2. Pipet out most of the old medium, but not completely (i.e., just enough to cover the surface of the well), from eachwell and very slowly along the wall of the well, add Medium N1(B) according to the following table.
Required volume per well
Reagents
Medium N1(B)

6-well plate24-well plate

96-well plate

2 ml

800 µl150 µl3. Incubate the cultures at 37°C, 5% CO2 for 2 days.
4. For subsequent medium changes, pipet out half (see volumes in the table above) of the old medium from each welland replace with an equal volume of room temperature Medium N1(B).
5. Repeat Step 4 every 2-3 days such as on Monday, Wednesday, and Friday for 1 week.

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