Ｏ-030 Dynamic culture on the microwell-patterned collagen scaffold supports the quality of isolated human islets

◎Kato Hiroyuki ¹ 　Chen Huajian² 　Izumi Kenji³ 　Koba Naoya³ 　Tsuchiya Takanori³ 　Kawazoe Naoki² 　Kandeel Fouad¹ 　Mullen Yoko¹ 　Chen Guoping² 　Komatsu Hirotake¹
シティーオブホープ¹ National_Institute_for_Materials_Science ² Tokai_Hit ³

Difficulty in maintaining isolated human islets prior to transplantation limits the clinical expansion of islet transplantations. In this study, we introduce a high seeding density culture platform for human islets by mimicking the physiological microenvironment including tissue fluidics and extracellular matrix support.
We cultured 3 human islet batches isolated from deceased donors for 2 week-culture using a dynamic culture system fabricated in a 35 mm-dish platforms. Within each dish, four 12 mm-diameter-cell culture inserts with porous polycarbonate membranes were embedded with silicone. The micropump-driven media circulation system (flow rate at 20 µL/min and 40 µL/min), including the 75 mL of media reservoir (replaced weekly), was integrated for the vertical media flow from the bottom to the top of the cell culture insert. Human islets (500 IEQ) were sandwiched between two layers of 200 µm-microwell-patterned microporous collagen scaffolds 12 mm in diameter and placed onto the cell culture insert. Static culture with 4 cell inserts using 75 mL of media in a 150-mm dish was prepared for controls. The entire setup was placed in a 5% CO2 incubator at 27C.
Dynamic cultures demonstrated significantly higher glutamine consumption when compared to the static controls. We observed a similar trend in glucose consumption but with no statistical significance. Islet viability was significantly higher in dynamic culture conditions compared to the controls. Islet morphology was well-maintained histologically in the dynamic culture conditions, whereas islets deteriorated in controls. No significant differences were seen between the two dynamic culture conditions. In summary, the physiological microenvironment-mimetic culture platform supported the quality of isolated human islets.