| | |
|---|
| Cell line: | RBL13 |
| DSMZ no.: | ACC 911 |
| Species: | human (Homo sapiens) |
| Cell type: | retinoblastoma |
| Origin: | established in 1986 from an unilateral retinoblastoma isolated from the enucleated eye of a 18 months old boy after pretreatment (type of treatment unknown); the RB1 gene is inactivated by promoter hypermethylation on two alleles and by a mutation on the third allele (Q62*) |
| Reference(s): | 40297, 40298, 40359 |
| Biosafety level: | 1 |
| Permissions and restrictions: | A |
| | DSMZ Cell Culture Data: |
| Morphology: | culture grows in spheroids of 1-3 mm in size; only after thawing as single cells and in small clumps, which aggregate into large spheroids; image; image; image |
| Medium: | 85% DMEM (with 4.5 g/l glucose) + 15% h.i. FBS + 10 µg/ml human insulin + 1x Glutamax + 1 mM sodium pyruvate + 50 µM mercaptoethanol |
| Subculture: | cells grow slowly and are difficult to culture; after thawing doubling time may be 2-3 weeks; cell 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 into one well in 4 ml medium; perform partial medium changes every 3 to 5 days; split 1:2 every 6-8 days by 2-3 times pipetting; to determine the number of cells: add Accutase for 5-10 minutes at 37°C and triturate 2x carefully; protocol |
| Incubation: | at 37 °C with 10% CO2 |
| Doubling time: | about 8-12 days |
| Harvest: | aggregates in one well of a split-ready 6-well may contain up to 6 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 MLPA (CpG106: 70%, CpG85: 40%); RB1 mutation status (exon 2 Q62*; Chr13:48307325C>T (hg38)) was verified by Sanger sequencing of all exons and flanking regions; heterozygosity was determined by microstallite analysis for chromosome 13 and copy number of 3 for chromosome 13 was confirmed by methylation-sensitive MLPA; it is assumed that two alleles are inactivated by DNA hypermethylation at the RB1 promoter and the unmethylated allele carries a nonsense mutation in RB1 resulting in a premature truncation |
| Viruses: | PCR: EBV -, HBV -, HCV -, HIV-1 -, HIV-2 -, HTLV-1/2 -, MLV -, 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 |
| | Print data sheet |
