Abstract:
BACKGROUND:Accurate islet quantification has proven difficult to standardize in a good manufacturing practices (GMP) approved manner. METHODS:The influence of assessment variables from both manual and computer-assisted digital image analysis (DIA) methods were compared using calibrated, standardized microspheres or islets alone. Additionally, a mixture of microspheres and exocrine tissue was used to evaluate the variability of both the current, internationally recognized, manual method and a novel GMP-friendly purity- and volume-based method (PV) evaluated by DIA in a semiclosed, culture bag system. RESULTS:Computer-assisted DIA recorded known microsphere size distribution and quantities accurately. By using DIA to evaluate islets, the interindividual manually evaluated percent coefficients of variation (CV%; n=14) were reduced by almost half for both islet equivalents (IEs; 31% vs. 17%, P=0.002) and purity (20% vs. 13%, P=0.033). The microsphere pool mixed with exocrine tissue did not differ from expected IE with either method. However, manual IE resulted in a total CV% of 44.3% and a range spanning 258 k IE, whereas PV resulted in CV% of 10.7% and range of 60 k IE. Purity CV% for each method were similar approximating 10.5% and differed from expected by +7% for the manual method and +3% for PV. CONCLUSION:The variability of standard counting methods for islet samples and clinical quantities of microspheres mixed with exocrine tissue were reduced with DIA. They were reduced even further by use of a semiclosed bag system compared with standard manual counting, thereby facilitating the standardization of islet evaluation according to GMP standards.
journal_name
Transplantationjournal_title
Transplantationauthors
Friberg AS,Brandhorst H,Buchwald P,Goto M,Ricordi C,Brandhorst D,Korsgren Odoi
10.1097/TP.0b013e31820ae48esubject
Has Abstractpub_date
2011-03-27 00:00:00pages
677-83issue
6eissn
0041-1337issn
1534-6080journal_volume
91pub_type
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