PUBLICATION
Comparison of tumor growth assessment using GFP fluorescence and DiI labeling in a zebrafish xenograft model
- Authors
- Dryer, Y., Berghausen, J., Creswell, K., Glasgow, E., Brelidze, T.I.
- ID
- ZDB-PUB-230717-40
- Date
- 2023
- Source
- Cancer biology & therapy 24: 22341402234140 (Journal)
- Registered Authors
- Glasgow, Eric
- Keywords
- Breast cancer, DiI, GFP, lipophilic dye, MDA-MB-231, xenograft, zebrafish
- MeSH Terms
-
- Animals
- Fluorescence
- Fluorescent Dyes/metabolism
- Green Fluorescent Proteins/genetics
- Heterografts
- Humans
- Neoplasms*
- Zebrafish*/metabolism
- PubMed
- 37455418 Full text @ Cancer Biol. Ther.
Citation
Dryer, Y., Berghausen, J., Creswell, K., Glasgow, E., Brelidze, T.I. (2023) Comparison of tumor growth assessment using GFP fluorescence and DiI labeling in a zebrafish xenograft model. Cancer biology & therapy. 24:22341402234140.
Abstract
DiI is a lipophilic fluorescent dye frequently used to label and trace cells in cell cultures and xenograft models. However, DiI can also transfer from labeled to unlabeled cells, including host organism cells, and label dead cells obscuring interpretation of the results. These limitations of DiI labeling in xenograft models have not been thoroughly investigated. Here we labeled green fluorescent protein (GFP)-expressing MDA-MB-231 cells with DiI to directly compare tumor growth assessment in zebrafish xenografts using the DiI labeling and GFP fluorescence. Our results indicate that the DiI based assessment significantly overestimated tumor growth in zebrafish xenograft models compared to the GFP fluorescence based assessment. The imaging of DiI labeled GFP-expressing MDA-MB-231 cell cultures indicated that the DiI labeling of the membrane is uneven. Analysis of the DiI labeled GFP-expressing MDA-MB-231 cell cultures with flow cytometry indicated that the DiI labeling varied over time while the GFP fluorescence remained unchanged, suggesting that the GFP fluorescence is a more reliable signal for monitoring tumor progression than the DiI labeling. Taken together, our results demonstrate limitations of using DiI labeling for xenograft models and emphasize the need for validating the results based on DiI labeling with other orthogonal methods, such as the ones utilizing genetically encoded fluorophores.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping