Isolation and characterization of a new cell line of retinoblastoma

BACKGROUNDː This article describes the characterization of a new cell line (IRB-1), which was established from an Iranian child with retinoblastoma.

METHODSː Using cell culture techniques, we isolated a clonal population from primary Cultures of retinoblastoma tumor samples and after several passages an established cell line, IRB-1, was obtained. The following techniques were performed: cell proliferation studies by MTT assays, scanning electron microscopy, cell surface analyses by flowcytometry, creation of neurospheres in soft agar and the expression of ABCG2, RAR-α, GFAP, and ACTB genes by real time PCR.

RESULTSː Of total 6 received samples, only a single cell line was successfully isolatedfrom a group E sporadic unilateral retinoblastoma. The primarily enucleated left eye had a differentiated retinoblastoma, with a slight degree of necrosis in pathology report. The adherent cultured cells were in spindle-shaped morphology with similar appendages and pulled neurons. Doubling time is 36 ± 5 hours. After 30 days, the effect of retinoic acid on isolated cells and expression of RAR-α receptor was appeared. Chromosomal abnormalities in chromosomes 3, 9, 10, 11, 14, 16, 17, and 22 were observed. Specific gene expression of ABCG2, RAR-α, and ACTB gene expression of GFAP positive expression of these genes in the control cells were observed. This cell line passaged 132 consecutive times.

Discussionː IRB-1 is a novel human adherent RB cell line, which with interesting characteristics of adherent and flocculent growth and more chromosomal imbalances different from other RB cell lines. A novel human RB cell line, IRB-1, was established from a 1-year old patient with a unilateral sporadic RB at Farabi Hospital- Tehran University Medical Sciences. The cultural characteristics of IRB-1 cell line were comparable to those of Y79 in characteristic.