User Guide
Quick-Neuron™ Excitatory - Maintenance Medium
Catalog Number: EX-MMIntroduction
Quick-Neuron™ Excitatory - Maintenance Medium may be used for the long-term maintenance of human pluripotent stem
cell-derived excitatory neurons following differentiation as outlined in the Quick-Neuron™ Excitatory - SeV Kit and Human
iPSC-derived Neurons user guides. Quick-Neuron™ Excitatory 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 theglutamatergic neuron marker vesicular glutamate transporter 1 (vGLUT1). When handled and maintained according to theinstructions in this user guide, excitatory neurons are viable long-term and are suitable for a variety of characterization andneurotoxicity assays.
Scale:The Quick-Neuron™ Excitatory - Maintenance Medium provides sufficient medium for 4 wells of a
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™ Excitatory - SeV Kit, Catalog Number: EX-SeV
Quick-Neuron™ Excitatory - Human iPSC-derived Neurons, Catalog Number: EX-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 N

840 μl

-20°C or -80°C

On ice or 4°C

Component G260 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 N(G2)1.

Prepare Medium N(G2) using the reagents listed in the table below.
○ Thaw each component 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 N(G2), and any subsequent media made with it, protected from light.
○ Store Medium N(G2) for up to 2 weeks at 4°C.
○ Leftover Components N and G2 can be discarded or saved at 4°C for up to two weeks.
Reagents

VolumeDMEM/F12

6 ml

Neurobasal Medium

6 ml

GlutaMAX

63 μl

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

126 μlComponent N1

391 μlComponent G2

12.6 μlFirst Week
1.

Prepare Medium N(G2P) 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.
○ Tapthe Component P tube 3 times and then briefly spin it down before use.
○ Store Medium N(G2P) for up to 2 weeks at 4°C.
○ Leftover Component P can be saved at 4°C.
Reagents

VolumeMedium N(G2)

5.2 mlComponent P2.6 μl

2. Pipet out half of the old medium from each well and very slowly along the wall of the well, add room temperature
Medium N(G2P) according to the following table.
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2Required volume per wellReagents
Medium N(G2P)

6-well plate24-well plate

96-well plate

1 ml

400 µl75 µl

3. Incubate the cultures at 37°C, 5% CO2 for 2 days.
4. Repeat Steps 2-3 every 2-3 days such as on Monday, Wednesday, and Friday for 1 week.

Second Week
1.

Warm Medium N(G2) 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 sampleimage below) users should continue using Medium N(G2P) in the second week, following the instructions toprepare Medium N(G2P) 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 N(G2) according to the following table.
Required volume per wellReagents
Medium N(G2)

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 N(G2).
5. Repeat Step 4 every 2-3 days such as on Monday, Wednesday, and Friday for 1 week.

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