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Cell line: | RBL20 |
DSMZ no.: | ACC 944 |
Species: | human (Homo sapiens) |
Cell type: | retinoblastoma |
Origin: | established in 1988 from a bilateral retinoblastoma isolated from the malignant primary tumor of the eye of a 16-month-old girl;
the RB1 gene is inactivated by splice site mutation in intron 6 (IVS6+1G>T) and loss of the wildtype allele |
Reference(s): | 40298, 40359, 40300, 40297 |
Biosafety level: | 1 |
Permissions and restrictions: | A |
| DSMZ Cell Culture Data: |
Morphology: | cells grow in suspension; after thawing as single cells and in small clumps; later with formation of three-dimensional aggregates (spheroids); image; image; image |
Medium: | 85% DMEM (with 4.5 g/l glucose) + 15% h.i. FBS + 10 µg/ml human insulin + 2 mM L-glutamine + 1 mM sodium pyruvate + 50 µM mercaptoethanol |
Subculture: | cells are difficult to culture; aggregates are sensitive to destruction by pipetting; use pipettes with wide opening (serological, wide-bore or cut); cultivate in 6-well plates; seed out all cells (approx 3 x 106 cells) into 1 well in 4 ml medium; perform partial medium changes every 3 to 5 days; when aggregates reach a size of 0.3 - 0.8 mm, split 1:2 to 1:3 once a week by 2-3 times pipetting; protocol |
Incubation: | at 37 °C with 10% CO2 |
Doubling time: | after thawing about 1-2 weeks, later 5-8 days |
Harvest: | aggregates in one well of a split-ready 6-well plate may contain up to 3-5 x 106 cells |
Storage: | frozen with 70% medium, 20% FBS, 10% DMSO |
| DSMZ Scientific Data: |
Mycoplasma: | negative in PCR assay |
Authenticity: | STR analysis according to the global standard ANSI/ATCC ASN-0002.1-2021 (2021) resulted in an authentic STR profile of the reference STR database. |
Molec. Genetics: | Methylation status of the RB1 promoter (CpG106) and differentially methylated region CpG85 (RB1 intron 2) was determined by methylation-senstive PCR (CpG106: 0%, CpG85: 100%); RB1 mutation status (splice site exon 6_IVS+1G>T; chr13:48349024G>T (hg38)) was verified by Sanger sequencing of all exons and flanking regions; loss of heterozygosity was determined by microstallite analysis for chromosome 13 and copy number of 1 for chromosome 13 was confirmed by methylation-sensitive MLPA |
Viruses: | PCR: EBV -, HBV -, HCV -, HIV-1 -, HIV-2 -, HTLV-1/2 -, MLV - |
Supplied as: | Delivery form | | Prices | | Frozen culture | | 900,- € | Growing culture (please inquire for exact delivery time) | | 1800,- € | DNA isolated from cell line (25 µg) | | 1120,- € | DNA isolated from cell line (5 µg) | | 280,- € | see price list |
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