RBL7

human (Homo sapiens)

retinoblastoma

established in 1987 from a bilateral retinoblastoma of a 16-month old boy after pretreatment (type of pretreatment unknown)

1

A

cells grow in irregular spheroids in suspension; after thawing as single cells and later in aggregates up to 1-2mm in size; image; image; image; image

85% DMEM (with 4.5 g/l glucose) + 15% h.i. FBS + 10 µg/ml human insulin + 2 mM L-glutamax + 1 mM sodium pyruvate + 50 µM mercaptoethanol

seed out all cells (approx 6-8 x 10⁶ cells) into 1 well of a 6-well plate with about 4 ml medium; split 1:2 every 5-8 days, by transferring 30-50% of the volume to a new well (take care not to disturb the spheroids: use pipettes with wide opening (serological, wide-bore or cut)) and work slowly and carefully; for determing cell number add Accutase for 5-10 min and triturate 2x carefully; cells will reaggregate thereafter; protocol

at 37 °C with 10% CO₂

about 8-12 days

aggregates in one well of a split-ready 6-well plate may contain up to 8 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 wildtype sequence was verified by Sanger sequencing of all exons and flanking regions; 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 -, HPV -, 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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