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Identification of stemness in primary retinoblastoma cells by analysis of stem-cell phenotypes and tumorigenicity with culture and xenograft models.
- Source :
-
Experimental Cell Research . Jun2019, Vol. 379 Issue 1, p110-118. 9p. - Publication Year :
- 2019
-
Abstract
- Retinoblastoma (RB) is a primary intraocular malignancy in childhood, and may develop relapse and metastatic disease. This study was to identify the stem-cell properties of primary retinoblastoma cells critical to tumorigenesis and metastasis. Primary cells were isolated from fresh human RB tissues after enucleation, and cultured in serum-free or serum-enriched conditions, with two RB cell-lines Weri-RB1 and Y79 for comparison. Proliferation of primary RB cells were well-maintained in serum-free condition of DMEM/F12 medium, and formed stem-cell like spheroids. The immaturity of cultured primary RB cells was demonstrated by tendency of highly expressed stem-cell markers (CD133, Nestin and OCT4) and suppressed mature retinal-cell markers (GFAP, MAP2 and Recoverin). CD133, a neural stem-cell marker being exclusively studied in RB, was found positive in small patches of cells in archival human RB by immunohistochemistry. Meanwhile, at initial isolation, insignificant CD133+ cells were detected by flow-cytometry, and substantial increase of positivity was observed after several days cultivated in serum-free condition. Cultured primary RB cells were engrafted in subretinal region of BALB/c nude mice for assessment of tumorigenicity. Strong tumorigenic activity and extensive progression of the xenograft retinoblastoma was induced by primary cells as compared with the two cell-lines. Again, immunohistochemistry confirmed that the stem-cell markers were emphasized in the xenograft tumor in mice. Our findings demonstrated that in comparison to the well-established RB cell-lines, cultured primary RB cells possess stem-cell like properties with highly expressed stem-cell markers, self-regenerative growth in culture, and strong in vivo oncogenic potentiality. Image 1 • Primary retinoblastoma (RB) cells were distinctive from RB cell-lines in-vitro. • Primary RB cells formed spheroids in serum-free culture conditions. • Primitivity of primary RB cells was shown by high expression of stem-cell markers. • Xenograft showed good oncogenicity of primary RB cells. [ABSTRACT FROM AUTHOR]
- Subjects :
- *RETINOBLASTOMA
*CANCER cell culture
*XENOGRAFTS
*PHENOTYPES
*CHILDHOOD cancer
Subjects
Details
- Language :
- English
- ISSN :
- 00144827
- Volume :
- 379
- Issue :
- 1
- Database :
- Academic Search Index
- Journal :
- Experimental Cell Research
- Publication Type :
- Academic Journal
- Accession number :
- 136342473
- Full Text :
- https://doi.org/10.1016/j.yexcr.2019.03.034