Development of quality control preparations for immunocytochemical assessment of urokinase-type plasminogen activator

Area of Science:

• Biochemistry
• Immunology
• Cell Biology

Background:

• Reproducibility and reliability are crucial for immunochemical and immunocytochemical analyses.
• Stable reference preparations are needed for quality assessment of antigen expression, including the plasminogen activation system.

Purpose of the Study:

• To develop quality control preparations for the immunocytochemical assessment of urokinase-type plasminogen activator (uPA).
• To establish a simple model system for creating reliable quality control materials for immunocytochemical assays.

Main Methods:

• Utilized cultured cell lines (BLM and IF6) with well-defined uPA expression levels, quantified by enzyme-linked immunosorbent assay (ELISA).
• Prepared cytospins and frozen sections from sucrose/Tissue-Tek blocks containing these cell lines.
• Evaluated the stability and homogeneity of uPA expression in the prepared quality control materials.

Main Results:

• Demonstrated stable and homogeneous urokinase-type plasminogen activator (uPA) expression in cytospins and frozen sections.
• Observed an excellent correlation between immunocytochemical staining results and ELISA-quantified uPA levels.
• Confirmed the availability of cell lines in large quantities for long-term production of identical quality control preparations.

Conclusions:

• Cell line-based cytospins or tissue blocks serve as a simple and effective system for establishing quality control preparations.
• This method ensures reproducible and reliable immunocytochemical assessment of urokinase-type plasminogen activator (uPA) and potentially other components of the plasminogen activation system.