Immune Therapeutic Applications

Cell therapy is undoubtedly a new hope to lead the future of biomedicine, but the application of human cells in medicine is not a new concept.

In the past few decades, cell therapy has made great progress, and cell therapy itself is no longer a simple collection of cells and infused back. Cells are now often required to be bioengineered, such as CAR-T cell therapy. We aim to provide you with standardized, GMP level equipment for cell quality control.

The Countstar product has been accepted by many companies leading the cell therapy, we can help our customer to build a stable, reliable cell concentration, viability monitor system.

Table of Content

• Challenge in Cell count and viability
• Dual Fluorescence viability counting by Countstar Rigel S2
• Determination of T/NK Cell Mediated Cytotoxicity
• Consistent Cell Counting and Global Data Management

Challenge in Cell count and viability
During all steps of clinical CAR-T cell manufacturing, the viability and cell count has to be determined precisely.Freshly isolated primary cells or cultured cells may contain impurities, several cell types or interfering particles such as cell debris which will make it impossible to analyze the cells of interest.

Dual Fluorescence cell count by Countstar Rigel S2
Acridine orange (AO) and Propidium iodide (PI) are nuclear nucleic acid binding dyes. AO can penetrate to both dead and live cells and stains the nucleated cells to generate green fluorescence. PI can stain the dead nucleated cells with compromised membranes and generate red fluorescence. The analysis excludes cell fragments, debris and artifacts particles as well as undersized events such as platelets, giving a highly accurate result. In conclusion, the Countstar S2 system can be used for every step of the cell manufacturing process.

A: AO/PI method can accurate distinguish the live and dead state of cells, and also can exclude the interference. By testing the diluting samples, dual-fluorescence method shows stable results.

Determination of T/NK Cell Mediated Cytotoxicity

By labeling the target tumor cells with non-toxic, non-radioactive calcein AM or transfect with GFP, we can monitor the killing of the tumor cells by CAR-T cells. While live target cancer cells will be labeled by a green calcein AM or GFP, the dead cells cannot retain the green dye. Hoechst 33342 is used for stain all cells (both T cells and tumor cells), alternatively, target tumor cells can be stained with membrane bound calcein AM, PI is used for stain the dead cells (both T cells and tumor cells). This staining strategy allows for the discriminate of different cells.

Consistent Cell Counting and Global Data Management
A common problem in conventional cell counting is the data differences between users, departments andsites. All Countstar analyzer counts the same in different location or production site. This is because in the process of quality control, each instrument must be calibrated to the standard instrument.

Central data bank allow user to keep all the data, like instrument test report, cell sample report and tester e-signature, safe and permanent.