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Immortalized-Human-Dopaminergic-Neuronal-Precursor-Cells-(LUHMES)

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Immortalized-Human-Dopaminergic-Neuronal-Precursor-Cells-(LUHMES)

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Immortalized-Human-Dopaminergic-Neuronal-Precursor-Cells-(LUHMES)

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	Print Version
BioSafety Level	II
Organism	Homo sapiens
Source Organ	8-week-old fetal human ventral mesencephalon
Donor Age	8 week
Donor Gender	undetermined
Growth Properties	Adherent
Morphology	Neuronal
Population Doubling	Subculture every 3-4 days
Recommended Seeding Density	150,000 cells/cm^2
Markers	DAT, VMAT-2, TH, α-SYN, β-III tubulin
Applications	For Research Use Only
Immortalization Method	Conditional immortalization by tetracyclin-controlled transduction with retrovirus carrying v-myc genes
Cell Type Characterization
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.
Procedure Overview
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Propagation	Use of PriCoat? T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. Grow the cells in NunclonTM culture vessel pre-coated with 50 μg/ml poly-L-ornithine (PLO) and 1 μg/ml fibronectin in H2O for at least 3 hours at 37°C. The base medium for this cell line is Advanced DME/F12 (Gibco; 12634010). To make the complete growth medium, add the following components to the base medium: 1x N2 supplement, 2 mM L-glutamine and 40 ng/ml recombinant bFGF. Atmosphere: air, 95%; Carbon dioxide (CO2), 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 the basal is Advanced DME/F12 (Gibco;12634010): 1x N2 supplement, 2 mM L-glutamine, 1mM dbcAMP, 1 μg/ml tetracycline, and 2 ng/ml recombinant human GDNF. Allow the cells in differentiation medium for 4-6 days before testing for neuron specific markers. Recommend to subculture when cells are grown to a cell density of 40-50%.
Quality Control	mRNA and protein expression levels of various markers pre and post differentiation are verified by RT-PCR and western blotting.
Disclaimer	1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. 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 strongly 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.