Application of Digital Image Analysis to Determine Pancreatic Islet Mass and Purity in Clinical Islet Isolation and Transplantation

摘要

Pancreatic islet mass, represented by islet equivalent (IEQ), is the most important parameter in decision making for clinical islet transplantation. To obtain IEQ, the sample of islets is routinely counted manually under a microscope and discarded thereafter. Islet purity, another parameter in islet processing, is routinely acquired by estimation only.In this study, we validated our digital image analysis (DIA) system developed using the software of Image Pro Plus for islet mass and purity assessment. Application of the DIA allows to better comply with current good manufacturing practice (cGMP) standards. Human islet samples were captured as calibrated digital images for the permanent record. Five trained technicians participated in determination of IEQ and purity by manual counting method and DIA. IEQ count showed statistically significant correlations between the manual method and DIA in all sample comparisons (r >0.819 and p < 0.0001). Statistically significant difference in IEQ between both methods was found only in High purity 100μL sample group (p = 0.029). As far as purity determination, statistically significant differences between manual assessment and DIA measurement was found in High and Low purity 100μL samples (p<0.005), In addition, islet particle number (IPN) and the IEQ/IPN ratio did not differ statistically between manual counting method and DIA.In conclusion, the DIA used in this study is a reliable technique in determination of IEQ and purity. Islet sample preserved as a digital image and results produced by DIA can be permanently stored for verification, technical training and islet information exchange between different islet centers. Therefore, DIA complies better with cGMP requirements than the manual counting method. We propose DIA as a quality control tool to supplement the established standard manual method for islets counting and purity estimation.