User Guide
Quick-Neuron™ Dopaminergic - Maintenance Medium
Catalog Number: DA-MMIntroduction
Quick-Neuron™ Dopaminergic - Maintenance Medium may be used for the long-term maintenance of human pluripotent
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 typicalneurite outgrowth and express a variety of neuronal markers, such as the pan-neuronal marker tubulin beta 3 class III(TUBB3) and the dopaminergic markers tyrosine hydroxylase (TH) and dopamine (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 ofcharacterization and neurotoxicity assays.
Scale:The Quick-Neuron™ Dopaminergic - Maintenance Medium provides sufficient medium for 4 wellsof 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™ Dopaminergic - mRNA Kit, Catalog Number: DA-mRNA
Quick-Neuron™ Dopaminergic - Human iPSC-derived Neurons, Catalog Number: DA-mRNA-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 D420 μl-20°C or -80°C

On ice or 4°C

Component D638 μl-20°C or -80°C

Room temperature

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(D4D6)
1.

Prepare Medium N(D4D6) 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(D4D6), and any subsequent media made with it, protected from light.
○ Store Medium N(D4D6) for up to 2 weeks at 4°C.
○ Leftover Components N, D4, and D6 can be discarded or saved at 4°C for up to two weeks.
Reagents

VolumeDMEM/F126 ml

Neurobasal Medium6 ml

GlutaMAX

63 μl

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

126 μlComponent N

391 μlComponent D4

12.6 μl

Component D6

12.6 μlFirst Week
1.

Prepare Medium N(D4D6P) 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 all tubes down before use.
○ Store Medium N(D4D6P) for up to 2 weeks at 4°C.
○ Leftover Component P can be saved at 4°C.
Reagents

VolumeMedium N(D4D6)

5.2 mlComponent P

5.2 μlelixirgensci.com

2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(D4D6P) according to the following table.
Required volume per well
Reagents
Medium N(D4D6P)

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(D4D6) 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 N(D4D6P) in the second week, following the instructions toprepare Medium N(D4D6P) 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(D4D6) according to the following table.
Required volume per well
Reagents
Medium N(D4D6)

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(D4D6).
5. Repeat Step 4 every 2-3 days such as on Monday, Wednesday, and Friday for 1 week.

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