RRC ID 73694
Author Nair RM, Revu NVL, Gali S, Kallamadi PR, Prabhu V, Manukonda R, Nemani H, Kaliki S, Vemuganti GK.
Title A short-term chick embryo in vivo xenograft model to study retinoblastoma cancer stem cells.
Journal Indian J Ophthalmol
Abstract PURPOSE:Cancer stem cells (CSCs) reported in various tumors play a crucial role in tumorigenesis and metastasis of retinoblastoma (Rb). Following the efforts to reduce, replace, and refine the use of mammalian models, we aimed to establish a short-term xenograft for Rb to evaluate the CSC properties of CD133- Rb Y79 cells, using the well-established chick embryo chorioallantoic membrane (CE-CAM) assay.
METHODS:Y79 cells were cultured, labeled with two different dyes (CM-Dil Y79 and enhanced green fluorescent protein (eGFP)) and sorted for CD133- and CD133 + subsets. Two million cells from each of the labeled groups were transplanted onto the abraded CAM on embryonic day 7 (E7). On E14, the tumor nodule formation on CAM and spontaneous metastasis to the embryos were evaluated by confocal microscopy, in vivo imaging, and histology.
RESULTS:Y79 cells formed pink-white raised perivascular nodules with feeder vessels on the CAM with both the types of labeled CD133- cells. CD133- cells, when compared to CD133 + cells, demonstrated significantly larger tumor volume (40.45 ± 7.744 mm3 vs 3.478 ± 0.69 mm3, P = 0.0014) and higher fluorescence intensity (CM-Dil: AUF = 6.37 × 107 ± 7.7 × 106 vs 1.08 × 107 ± 1.6 × 106; P < 0.0001; eGFP: AUF = 13.94 × 104 ± 2.54 × 104 vs AUF = 1.39 × 104 ± 0.4 × 104; P = 0.0003). The metastatic potential of CD133- cells was also observed to be higher as noted by in vivo imaging and histopathology.
CONCLUSION:This study highlights that CE-CAM is a feasible alternative nonmammalian model for evaluating tumorigenicity and metastatic potential of Y79 CSCs. Increased tumorigenicity and metastatic potential of CD133- subset of tumor cells substantiate their CSC properties.
Volume 70(5)
Pages 1703-1711
Published 2022-5-1
DOI 10.4103/ijo.IJO_2348_21
PII IndianJOphthalmol_2022_70_5_1703_343968
PMID 35502056
PMC PMC9332960
MeSH AC133 Antigen / metabolism Animals Cell Line, Tumor Chick Embryo Heterografts Humans Mammals / metabolism Neoplastic Stem Cells / metabolism Neoplastic Stem Cells / pathology Retinal Neoplasms* / pathology Retinoblastoma* / pathology
Human and Animal Cells Y79(RCB1645)