RBL30

human (Homo sapiens)

retinoblastoma

established in 1988 from an unilateral retinoblastoma of a 30-month old boy after pretreatment (type of pretreatment unknown)

40298, 40359, 40297

1

A

cells grow in suspension; after thawing as single cells and in small clumps; later with formation of three-dimensional aggregates (spheroids) up to 5mm; image; image; image

85% DMEM (with 4.5 g/l glucose) + 15% h.i. FBS + 10 µg/ml human insulin + 4 mM L-glutamine + 1 mM sodium pyruvate + 50 µM mercaptoethanol

seed out all cells (approx 6 x 10⁶ cells) into 1 well of 6-well plate; split 1:2 every 5-6 days, by transferring 50% of the volume to a new well (take care not to disturb the spheroids: use pipettes with wide opening (serological, wide-bore or cut)); for determing cell number add Accutase for 5-10 min and triturate 2x carefully protocol

at 37 °C with 10% CO₂

ca. 5 days

aggregates in one well of a split-ready 6-well plate may contain up to 6 x 10⁶ cells

frozen with 70% medium, 20% FBS, 10% DMSO

negative in PCR assay

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.

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 (exon 8 R251*; chr 13:48362847C>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 2 for chromosome 13 was confirmed by methylation- sensitive MLPA

PCR: EBV -, HBV -, HCV -, HIV-1 -, HIV-2 -, HTLV-1/2 -, MLV -

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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