abm | Immortalized Human Dopaminergic Neuronal Precursor Cells (LUHMES) | T0284

abm | Immortalized Human Dopaminergic Neuronal Precursor Cells (LUHMES) | T0284

Immortalization Method:

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

BioSafety Level:

II

Organism:

Homo sapiens

Species:

Human

Source Organ:

8-week-old fetal human ventral mesencephalon

Organ Type:

Brain

Growth Properties:

Adherent

Morphology:

Flattened|Dendritic processes

Passage Number:

N/A

Population Doubling:

30 - 40 hours

Seeding Density:

50,000 - 100,000 cells/cm10²; cells will have viability of around 40-50%. Post thawing, it will need 4-5 days for cells to recover in culture; Recommended split ratio is 1:4 to 1:5

Markers:

DAT, VMAT-2, TH, α-SYN, β-III tubulin

Donor Age:

8 week

Donor Gender:

undetermined

Donor Ethnicity:

N/A

Propagation:

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.
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: 1x N2 supplement (ThermoFisher Scientific), 2 mM L-glutamine and 40 ng/ml recombinant bFGF (Z101455).
Carbon dioxide (CO₂): 5%, Temperature: 37.0°C.

Quality Control:

mRNA and protein expression levels of various markers pre and post differentiation are verified by RT-PCR and western blotting.

Shipping Condition:

Dry Ice

Storage Condition:

liquid nitrogen or -180C