Brain-tropic metastatic derivatives (BrM) of cell lines MDA231, HCC1954, and MMTV-ErbB2 were established as previously described11 (link),12 (link),94 (link). BrM derivatives of cell line E0771 were generated following the same procedure via two cycles of in vivo selection of the parental cells for their preferential metastasis to the brain in background-matched FVB/NJ mice (The Jackson Laboratory, strain #001800). Briefly, E0771 cells were transduced with lentivirus expressing the triple-fusion reporter encoding herpes simplex virus thymidine kinase 1 (HSV1-TK), GFP and firefly luciferase96 (link). 1.0 × 105 parental cells were injected into the left ventricle of anesthetized 5–7 week-old FVB/NJ mice (The Jackson Laboratory, strain #: 001800) in a volume of 100 μL. Tumor development was monitored by weekly bioluminescence imaging (BLI) using the Xenogen IVIS-200 system (PerkinElmer). We harvested and dissociated brains with positive BLI signal into single-cell suspension, and isolated GFP+ cells by FACS sorting to be subject to a second round of in vivo selection to obtain BrM derivatives.
Protocol detail
Establishment of Brain-tropic Metastatic Cell Lines
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Brains
Cell
Cell line
Derivatives
Erbb2
Firefly
Herpes simplex virus 1
Left ventricle
Lentivirus
Metastasis to brain
Mice
Mmtv
Parental
Strain
Thymidine kinase
Tumor
Corresponding Organization : Memorial Sloan Kettering Cancer Center
Other organizations : Westlake University, Howard Hughes Medical Institute
Top 5 similar protocols
Variable analysis
independent variables
- Lentiviral transduction of E0771 cells with triple-fusion reporter encoding HSV1-TK, GFP, and firefly luciferase
- Injection of transduced E0771 cells into the left ventricle of anesthetized 5-7 week-old FVB/NJ mice
- Two cycles of in vivo selection for preferential metastasis to the brain
- Tumor development monitored by weekly bioluminescence imaging (BLI)
- Isolation of GFP+ cells from brain tissue with positive BLI signal
- Background-matched FVB/NJ mice (The Jackson Laboratory, strain #001800)
- Positive control: Parental E0771 cells transduced with the triple-fusion reporter
- Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!