IRES-GFP Stable LUHMES Cell Line

CAT.NOUNITPRICE
T64571x106 cells / 1.0 ml
$0.00

Specifications


Description

Through lentiviral infection, the IRES-GFP Stable LUHMES Cell Line was generated.
abm also offers A30P aSYN-IRES-GFP Stable LUHMES Cell Line (T6454), and A53T aSYN-IRES-GFP Stable LUHMES Cell Line (T6455), Wild Type aSYN-IRES-GFP Stable LUHMES Cell Line (T6456).

SKUT6457
SpeciesHuman (H. sapiens)
Tissue/Organ/Organ SystemBrain
Growth PropertiesAdherent
Cell MorphologyEpithelial
Applications

For Research Use Only

Unit quantity1x106 cells / 1.0 ml
Cell TypeDrug Discovery Cell Lines
Expression Profile

GFP, Tetracycline resistance

Propagation Requirements

Grow the cells in culture vessel pre-coated with 50 μg/ml poly-L-ornithine (PLO) (TM062) and 1 μg/ml fibronectin (EMD Millipore; Cat. FC010) in H2O for at least 3 hours at 37°C. Do not grow these cells in culture vessels with surface areas equal to less than 12.5 cm2. These cells do not grow in 6-well, 24-well, 48-well, or 96-well plates.

The base medium for this cell line is Advanced DMEM/F12 (Gibco;12634010). To make the complete growth medium, add the following components to the base medium: N2 supplement (ThermoFisher Scientific) to a final concentration of 1X, L-glutamine (G275) to a final concentration of 2 mM, and Recombinant Human FGF2 (Z101455) to a final concentration of 40 ng/ml. Cells may be grown in the presence of 1 μg/ml Tetracycline.
Change media every 2-3 days. Do not let media colour change to orange-yellow.
The cells will form round clumps instead of a monolayer when stressed. It is recommended to subculture when cells are grown to a cell density of 50-60%.
Carbon dioxide (CO2): 5%, Temperature: 37.0°C.
To subculture the cells, use 0.25% Trypsin-EDTA (TM051). After adding trypsin, incubate the cells at room temperature for 2-3 minutes and agitate the culture vessel until 90% of the cells have detached. Immediately neutralize the trypsin using Trypsin Neutralizing Solution (Lonza, Cat. CC-5002). Add Trypsin Neutralizing Solution twice the volume of trypsin used. Centrifuge at 200x g for 2-3 minutes. Aspirate supernatant and resuspend cells in complete media. Plate cells into pre-warmed PLO-fibronectin-coated vessels at 37.0°C. Avoid subculturing if the cells appear stressed. Note: If leaving the cells over the weekend (or more than 2 days), make sure to do a high split ratio (1:4 to 1:5).

To differentiate the cells into neurons, change the medium to differentiation medium after the cells have grown to a density of 40-50%.
To make complete differentiation medium, add the following components to Advanced DMEM/F12 (Gibco;12634010): N2 supplement to a final concentration of 1X, L-glutamine (G275) to a final concentration of 2 mM, dbcAMP to a final concentration of 1 mM, tetracycline to a final concentration of 1 μg/ml, and Recombinant Human GDNF (Z101055) to a final concentration of 2 ng/ml. Allow the cells to grow in differentiation medium for 4-6 days before testing for neuron specific markers.

QC

1) Western blot; 2) Immunocytochemistry

Disclaimer

1. For for-profit organizations and corporations, please contact [email protected] for pricing of this item.

2. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected].

3. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.

4. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).

5. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information.

6. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.

7. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."

DepositorGeorg-August-Universität Göttingen
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