|
In vitro assays
|
Immunophenotyping assay
|
ratio of CD4 and CD8
|
Easy and quick
|
Cannot be used a predictive biomarker due to variations from each individual.
|
|
Cannot reflect the phenotyping in vivo
|
|
Proliferation and Cytokine secretion assay
|
IL-2 and IFN-gamma productions
|
Easy and quick
|
Cannot reflect the proliferation and cytokine productions in vivo
|
|
Cytotoxicity by standard 51Cr-release assay
|
4-hour killing of tumor cells
|
Easy and quick
|
Short -term in vitro activation
|
|
No interaction with host real tumor cells
|
|
Long-term killing assay
|
Number of CAR-T cells
|
Reflects the CAR-T expansion and tumor killing in 1- or 2-week in vitro assay
|
Time-consuming
|
|
Number of artificial tumor cells
|
Artificial modified tumor cell lines
|
|
Slow and variable
|
|
Technically complex
|
|
Available in vitro Strategies for clinical use CAR-T cells
|
Vector copy number (VCN) and CAR expression
|
Technically convenient
|
Cannot be used as predictive biomarker due to variations from each individual.
|
|
In vivo assays
|
NSG mouse model
|
Tumor size and tumor growth
|
Predicts persistence of CAR cells
|
No host immune system
|
|
Mouse survival
|
Predicts CAR-T killing capability
|
No tumor microenvironment (TME)
|
|
Body weight
|
Easy to use
|
Expensive
|
|
Labor-intensive
|
|
Syngeneic transplantable model
|
Tumor size
|
Intact host immune system
|
Slow and complex
|
|
Mouse survival
|
Some TME development
|
Model is variable
|
|
Body weight
|
Expensive
|
|
Biodistribution of CAR cells
|
Labor-intensive
|
