›› 2018, Vol. 38 ›› Issue (8): 865-.doi: 10.3969/j.issn.1674-8115.2018.08.003

• Original article (Basic research) • Previous Articles     Next Articles

Establishment and genomic characterization of a novel retinoblastoma cell line without RB1 mutation derived a Han Chinese donor

YUAN Xiao-ling1, 2, HE Xiao-yu1, 2, LI Yong-yun1, 2, CHAI Pei-wei1, 2, WANG Hui-xue1, 2, JIA Ruo-bing1, 2, XU Yang-fan1, 2, SHANG Qing-feng1, 2, ZHAO Hong-bo1, 2, JIA Ren-bing1, 2   

  1. 1. Department of Ophthalmology, Shanghai Ninth Peoples Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China; 2. Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology, Shanghai 200011, China
  • Online:2018-08-28 Published:2018-09-11
  • Supported by:
    Scientific Research Program for Public Welfare Industry of Ministry of Health of China, 201402014; National Natural Science Foundation of China, 81372902; Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Grant, 20152223; Shanghai Municipal Key Laboratory, 17DZ2260100

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Abstract: Objective · To develop and characterize a novel retinoblastoma cell line a Han Chinese patient. Methods · Cells were dispersed tumor tissue harvested an enucleated eyeball harbouring retinoblastoma under sterile conditions. The dissociated cells were cultured, purified and passaged in vitro. Morphologic and genetic analysis and detection of surface biomarkers were performed on the cell line and primary tumorimmunofluorescence, immunohistochemical staining, DNAshort tandemrepeat (STR) analysis, karyotype analysis, and exome sequencing. Results · This human retinoblastoma cell line was designated as SNPH-Rb-C24. It expressed neural cell adhesion molecule 1 and synaptophysin, which confirmed its neuronal derivation. DNA STR analysis showed an identical match between SNPH-Rb-C24 and primary tumor. Karyotype analysis showed complex chromosomal abnormalities in SNPH-Rb-C24, while no alteration in 13q was observed. Comparative exome sequencing identified common mutated genes and RB1+/+ in both SNPH-Rb-C24 and primary tumor. Orthotopic xenograft tumors derived early passage cells were established. Conclusion · A human retinoblastoma cell line (SNPH-Rb-C24) derived a Han Chinese patient with RB1+/+ retinoblastoma is developed, which retains critical biological and genomic features of the donor tumor.

Key words: retinoblastoma, primary cell line, RB1 gene, karyotypeanalysis, exome sequencing

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