User Guide

Quick-NeuronTM Dopaminergic - Maintenance Medium
Catalog Number: DA-MMIntroduction
Quick-Neuron™ Dopaminergic - Maintenance Medium may be used for the long-term maintenance of humanpluripotent stem cell-derived dopaminergic neurons following differentiation as outlined in the Quick-Neuron™
Dopaminergic - mRNA Kit and Human iPSC-derived Neurons user guides. Quick-Neuron™ Dopaminergic differentiated cell cultures display typical neurite outgrowth and express a variety of neuronal markers, such as the pan-neuronalmarker tubulin beta 3 class III (TUBB3) and the dopaminergic markers tyrosine hydroxylase (TH) anddopamine (DA). When handled and maintained according to the instructions in this user guide, dopaminergic neurons
are viable long-term and are suitable for a variety of characterization and neurotoxicity assays.
Scale: 					The Quick-Neuron™ Dopaminergic - Maintenance Medium provides sufficient medium for 4
wells of a 24-well plate for up to 2 weeks.

Related Products:			

Quick-Neuron™ Dopaminergic - mRNA Kit, Catalog Number: DA-mRNA
Quick-Neuron™ Dopaminergic - Human iPSC-derived Neurons, Catalog Number: DA-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 D420 μl-20°C or -80°C

Component D6

16 μl-20°C or -80°C

Component P

14 μl-20°C or -80°C

Required Consumables
ItemVendor

Catalog 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 mlNeurobasal Medium8 ml

200 mM Glutamax (100x)

83 μl

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

167 μlComponent N517 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(D4D6P)
1. Prepare Medium N(D4D6P) using the reagents listed in the table below.
• Thaw Component D4 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. Spin down Component D6before use.
Medium N(D4D6P) Reagents

VolumeMedium N

5.5 mlComponent D4

5.5 μlComponent D6

5.5 μlComponent P

2.8 μl2. Save the leftover Component D4 and Component D6 at 4°C.
• The leftover Component P can be discarded or saved for another use.
3. Warm Medium N(D4D6P) 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(D4D6P).
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.

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Second Week
Medium N(D4D6)
1. Prepare fresh Medium N(D4D6) using the reagents listed in the table below.
• Place Component D4 on ice. Spin down before use.
• Warm Component D6 at room temperature for 30 minutes. Spin down before use.
• Warm Medium N at room temperature for 30 minutes.
Medium N(D4D6) Reagents

VolumeMedium N

7 mlComponent D4

7 μl

Component D67 μl

2. Warm Medium N(D4D6) 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(D4D6) 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(D4D6).
6. Repeat Step 5 every 2-3 days such as on Monday, Wednesday, and Friday for 1 week.

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