A short-term chick embryo in vivo xenograft model to study retinoblastoma cancer stem cells
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 propertie...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wolters Kluwer Medknow Publications
2022-01-01
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| Series: | Indian Journal of Ophthalmology |
| Subjects: | |
| Online Access: | http://www.ijo.in/article.asp?issn=0301-4738;year=2022;volume=70;issue=5;spage=1703;epage=1711;aulast=Nair |
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| author | Rohini M Nair Narayana V L Revu Sucharita Gali Prathap Reddy Kallamadi Varsha Prabhu Radhika Manukonda Harishankar Nemani Swathi Kaliki Geeta K Vemuganti |
| author_facet | Rohini M Nair Narayana V L Revu Sucharita Gali Prathap Reddy Kallamadi Varsha Prabhu Radhika Manukonda Harishankar Nemani Swathi Kaliki Geeta K Vemuganti |
| author_sort | Rohini M Nair |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-12T12:39:36Z |
| format | Article |
| id | doaj.art-12ab43072ff04970b28aa42ad6103e76 |
| institution | Directory Open Access Journal |
| issn | 0301-4738 1998-3689 |
| language | English |
| last_indexed | 2024-12-12T12:39:36Z |
| publishDate | 2022-01-01 |
| publisher | Wolters Kluwer Medknow Publications |
| record_format | Article |
| series | Indian Journal of Ophthalmology |
| spelling | doaj.art-12ab43072ff04970b28aa42ad6103e762022-12-22T00:24:14ZengWolters Kluwer Medknow PublicationsIndian Journal of Ophthalmology0301-47381998-36892022-01-017051703171110.4103/ijo.IJO_2348_21A short-term chick embryo in vivo xenograft model to study retinoblastoma cancer stem cellsRohini M NairNarayana V L RevuSucharita GaliPrathap Reddy KallamadiVarsha PrabhuRadhika ManukondaHarishankar NemaniSwathi KalikiGeeta K VemugantiPurpose: 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.http://www.ijo.in/article.asp?issn=0301-4738;year=2022;volume=70;issue=5;spage=1703;epage=1711;aulast=Naircancer stem cellschick embryometastasisretinoblastomaxenograft model |
| spellingShingle | Rohini M Nair Narayana V L Revu Sucharita Gali Prathap Reddy Kallamadi Varsha Prabhu Radhika Manukonda Harishankar Nemani Swathi Kaliki Geeta K Vemuganti A short-term chick embryo in vivo xenograft model to study retinoblastoma cancer stem cells Indian Journal of Ophthalmology cancer stem cells chick embryo metastasis retinoblastoma xenograft model |
| title | A short-term chick embryo in vivo xenograft model to study retinoblastoma cancer stem cells |
| title_full | A short-term chick embryo in vivo xenograft model to study retinoblastoma cancer stem cells |
| title_fullStr | A short-term chick embryo in vivo xenograft model to study retinoblastoma cancer stem cells |
| title_full_unstemmed | A short-term chick embryo in vivo xenograft model to study retinoblastoma cancer stem cells |
| title_short | A short-term chick embryo in vivo xenograft model to study retinoblastoma cancer stem cells |
| title_sort | short term chick embryo in vivo xenograft model to study retinoblastoma cancer stem cells |
| topic | cancer stem cells chick embryo metastasis retinoblastoma xenograft model |
| url | http://www.ijo.in/article.asp?issn=0301-4738;year=2022;volume=70;issue=5;spage=1703;epage=1711;aulast=Nair |
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