User Guide

Quick-NeuronTM 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 andHuman iPSC-derived Neurons user guides. Quick-Neuron™ Excitatory 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 glutamatergic neuron marker vesicular glutamate transporter 1 (vGLUT1). When handled andmaintained according to the instructions in this user guide, excitatory neurons are viable long-term and are suitable for avariety of characterization and neurotoxicity assays.
Scale: 					The Quick-Neuron™ Excitatory - Maintenance Medium provides sufficient medium for 4		
wells of a 24-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-CWKit Contents
Upon receipt, store the reagents at the temperatures indicated in the table below. All reagents are shipped on dry ice.
Reagents

VolumeStorage

Component N

840 μl

-20°C or -80°C

Component G2

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

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

VolumeDMEM/F128 ml

Neurobasal Medium

8 ml

200 mM Glutamax (100x)

83 μl

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

167 μlComponent N

517 μl

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

First Week
Medium N(G2P)
1. Prepare Medium N(G2P) using the reagents listed in the table below.
• Thaw Component G2 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 N(G2P) Reagents

VolumeMedium N

5.5 mlComponent G25.5 μl

Component P

2.8 μl2. Save the leftover Component G2 at 4°C.
• The leftover Component P can be discarded or saved for another use.
3. Warm Medium N(G2P) at room temperature for 20-30 minutes until it no longer feels cold.
4. Pipet out half (400 μl) of the old medium from each well using a P1000 pipettor and add 400 μl Medium N(G2P).
5. Incubate the cultures at 37°C, 5% CO2 for 2 days.
6. Repeat Steps 3-5 every 2-3 days such as on Monday, Wednesday, and Friday for 1 week.

Second Week
Medium N(G2)
1. Prepare fresh Medium N(G2) using the reagents listed in the table below.
• Warm Medium N at room temperature for 30 minutes.
• Place Component G2 on ice. Spin down before use.
Medium N(G2) Reagents

VolumeMedium N

7 ml

Component G2

7 μl

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