Immortalized Human Dopaminergic Neuronal Precursor Cells (LUHMES)

Cat. No.
T0284
Unit
1x106 cells / 1.0 ml
Price
$440.00

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Cat. No. T0284
Name Immortalized Human Dopaminergic Neuronal Precursor Cells (LUHMES)
Description

The Immortalized Human Dopaminergic Neuronal Precursor Cells, also known as the Lund Human Mesencephalic (LUHMES) cell line, is a subclone of the tetracycline-controlled, v-myc-overexpressing human mesencephalic-derived cell line MESC2.10. This cell line is unique in that it can be differentiated to acquire a dopaminergic neuron-like phenotype under appropriate growth conditions. LUHMES expresses functional dopamine transporter (DAT), vesicular monoamine transporter (VMAT-2), tyrosine hydroxylase (TH) and the neuronal form of β-III tubulin after differentiation. In addition to retaining dopaminergic neuronal-specific markers, LUHMES also exhibit electrophysiological properties, thus making this cell line a valuable neuronal model for neurodevelopmental studies, disease modelling and neuropharmacology. May take 1.4 to 5 days for cells to recover in culture.

Organism Human (H. sapiens)
Tissue Brain
Donor History 8-week-old fetal human ventral mesencephalon
Growth Properties Adherent, flat dendritic processes
Cell Type Immortalized Cells
Unit 1x106 cells / 1.0 ml
Storage Condition Vapor phase of liquid nitrogen, or below -130°C.
Shipping Conditions Ship with dry ice.
Product Format Frozen
Intended Use This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
BioSafety II
Certificate of Analysis For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com.
Growth Conditions

Grow cells in culture vessels 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 cells in culture vessels with surface areas equal to or less than 12.5 cm²; cells do not grow in 6-well, 24-well, 48-well, or 96-well plates. Advanced DMEM/F12 (Gibco;12634010) + 1X N2 supplement (ThermoFisher Scientific) + 2 mM L-glutamine (G275) + 40 ng/ml rh FGF2 (Z101455) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂.  Do not let media colour change to orange-yellow. 

For information regarding cell differentiation, please refer to the Differentiation Protocol PDF under the Documents Tab. 

Unpacking and Storage Instructions

1. Visually examine the packaging containers for signs of leakage or breakage.

2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130°C, preferably in liquid nitrogen vapor phase storage, until ready for use.

To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability.


Thawing Protocol

1. Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.

2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.

3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.

4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.

5. Incubate the cells at the recommended conditions.

Subculture Protocol

Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent.

1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.

2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment.

3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.

4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.

5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired.

6. Incubate the cells at the recommended conditions.

Cryopreservation

Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.

Seeding Density (cells/cm2) 50,000 - 100,000
Split Ratio 1:4 to 1:5
Population Doubling Time (h) 30 - 40
Immortalization Method

Conditional immortalization by tetracyclin-controlled transduction with retrovirus carrying v-myc genes

Expression

DAT, VMAT-2, TH, α-SYN, β-III tubulin

STR Profiling

D5S818 : 11,13

D13S317 : 9,11

D7S820 : 11,13

D16S539 : 11,12

VWA : 14,17

TH01 : 7,9.3

AMEL : X,X

TPOX : 8,8

CSF1PO : 13,14

D12S391 : 22,22

FGA : 19,21

D2S1338 : 17,24

D21S11 : 30,31

D18S51 : 12,12

D8S1179 : 12,13

D3S1358 : 17,18

D6S1043 : 12,14

PENTAE : 11,13

D19S433 : 14,15

PENTAD : 12,13

D1S1656 : 14,15

Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0284.
Warranty 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”.
Disclaimer
1. 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.

2. 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).

3. 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.

4. 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.

5. 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."
Depositor University of Konstanz
Application Research Use Only.
Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0284
Print & Download Datasheet
  • Scholz, D., Pöltl, D., Genewsky, A., Weng, M., Waldmann, T., Schildknecht, S., & Leist, M. (2011). Rapid, complete and large-scale generation of post-mitotic neurons from the human LUHMES cell line. Journal of Neurochemistry, 119(5), 957–971. https://doi.org/10.1111/j.1471-4159.2011.07255.x

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