|Organism||Human (Homo sapiens), 77 years old, Male|
|Cell type||Glioma stem cell / Glioblastoma stem cells|
|Growth mode||Adherent / Sphere|
|Doubling time||Intermediate proliferative capacity|
|Tumorigenic||Yes (immunocompromised mice)|
|Related cell lines|
CELL LINE ISOLATION INFORMATION
The cell line U3065MG was established from a Glioblastoma tumor biopsy taken from a 77 years old male patient.
Dissociated cells from tumor tissue was allowed to grow as spheres onto uncoated dishes in complete culture medium [1:1 mix of Neurobasal and DMEM/F12 media supplemented with N2 (1x final conc), B27 (1x final conc), human recombinant FGF2 (final conc. 10 ng/ml) and human EGF (final conc. 10 ng/ml)]. These neurospheres were plated onto laminin coated dishes after 5 to 7 days to allow them to grow as adherent culture. Please read Xie et al., 2015 supplementary data for exact details.
Recommended propagation method
Culture medium: Neurobasal and DMEM/F12 media (1:1 mix) containing N2 and B27 supplements and human recombinant FGF2 and EGF (10 ng/ml)
Culture dishes: Primaria dishes coated with mouse laminin
Culture conditions: Humidified incubator, 95% air + 5% CO2, 37°C
Split ratio: 1:4 to 1:6
Split frequency: ?
Harvesting time: Subconfluent culture
Split Method: TrypLE
Recommended Cryopreservation Method
Freeze medium: Culture medium (complete medium containing all ingredients including FGF2 and EGF) supplemented with 10% DMSO
Storage temperature: Liquid nitrogen vapor phase / -150 °C freezer
Cell density: ≈ 5 x 105 cells/ml
- Xie et al., 2015. The Human Glioblastoma Cell Culture Resource: Validated Cell Models Representing All Molecular Subtypes. EBioMedicine. 2015 Aug 15;2(10):1351-63. PMID-26629530; Full Text Link: thelancet (open access), PMC