BACKGROUND: We evaluated three commercially available enzymes for pancreatic digestion by comparing key parameters during the islet isolation process, as well as islet quality post-isolation. METHODS: Retrospectively compared and analyzed islet isolations from pancreata using three different enzyme groups: Liberase HI (n=63), Collagenase NB1/Neutral Protease (NP) (n=43), and Liberase Mammalian Tissue Free Collagenase/Thermolysin (MTF C/T) (n=115). A standardized islet isolation and purification method was used. Islet quality assessment was carried out using islet count, viability, in vitro glucose-stimulated insulin secretion (GSIS), glucose-stimulated oxygen consumption rate (ΔOCR), and in vivo transplantation model in mice. RESULTS: Donor characteristics were not significantly different among the three enzyme groups used in terms of age, sex, hospital stay duration, cause of death, body mass index (BMI), hemoglobin A1c (HbA1c), cold ischemia time (CIT), and pancreas weight. Digestion efficacy (percentage of digested tissue by weight) was significantly higher in the Liberase MTF C/T group (73.5 ± 1.5 %) when compared to the Liberase HI group (63.6 ± 2.3 %) (p<0.001) and the Collagenase NB1/NP group (61.7 ± 2.9%) (p<0.001). The stimulation index for GSIS was significantly higher in the Liberase MTF C/T group (5.3 ± 0.5) as compared to the Liberase HI (2.9 ± 0.2) (p<0.0001) and the Collagenase NB1/NP (3.6 ± 2.9) (p=0.012) groups. Furthermore, the Liberase MTF C/T enzymes showed the highest success rate of transplantation in diabetic NOD Scid mice (65%), which was significantly higher than the Liberase HI (42%, p=0.001) and the Collagenase NB1/NP enzymes (41%, p<0.001). CONCLUSIONS: Liberase MTF C/T is superior to Liberase HI and Collagenase NB1/NP in terms of digestion efficacy and glucose-stimulated insulin secretion in vitro. Moreover, Liberase MTF C/T had a significantly higher success rate of transplantation in diabetic NOD Scid mice compared to Liberase HI and Collagenase NB1/NP enzymes.
BACKGROUND: We evaluated three commercially available enzymes for pancreatic digestion by comparing key parameters during the islet isolation process, as well as islet quality post-isolation. METHODS: Retrospectively compared and analyzed islet isolations from pancreata using three different enzyme groups: Liberase HI (n=63), Collagenase NB1/Neutral Protease (NP) (n=43), and Liberase Mammalian Tissue Free Collagenase/Thermolysin (MTF C/T) (n=115). A standardized islet isolation and purification method was used. Islet quality assessment was carried out using islet count, viability, in vitro glucose-stimulated insulin secretion (GSIS), glucose-stimulated oxygen consumption rate (ΔOCR), and in vivo transplantation model in mice. RESULTS:Donor characteristics were not significantly different among the three enzyme groups used in terms of age, sex, hospital stay duration, cause of death, body mass index (BMI), hemoglobin A1c (HbA1c), cold ischemia time (CIT), and pancreas weight. Digestion efficacy (percentage of digested tissue by weight) was significantly higher in the Liberase MTF C/T group (73.5 ± 1.5 %) when compared to the Liberase HI group (63.6 ± 2.3 %) (p<0.001) and the Collagenase NB1/NP group (61.7 ± 2.9%) (p<0.001). The stimulation index for GSIS was significantly higher in the Liberase MTF C/T group (5.3 ± 0.5) as compared to the Liberase HI (2.9 ± 0.2) (p<0.0001) and the Collagenase NB1/NP (3.6 ± 2.9) (p=0.012) groups. Furthermore, the Liberase MTF C/T enzymes showed the highest success rate of transplantation in diabeticNOD Scid mice (65%), which was significantly higher than the Liberase HI (42%, p=0.001) and the Collagenase NB1/NP enzymes (41%, p<0.001). CONCLUSIONS: Liberase MTF C/T is superior to Liberase HI and Collagenase NB1/NP in terms of digestion efficacy and glucose-stimulated insulin secretion in vitro. Moreover, Liberase MTF C/T had a significantly higher success rate of transplantation in diabeticNOD Scid mice compared to Liberase HI and Collagenase NB1/NP enzymes.
Authors: T J Fetterhoff; T J Cavanagh; K J Wile; M J Wright; F E Dwulet; J Gill; B Ellis; M E Smith; J K Critser; M Zieger Journal: Transplant Proc Date: 1995-12 Impact factor: 1.066
Authors: J F Gill; L L Chambers; J L Baurley; B B Ellis; T J Cavanaugh; T J Fetterhoff; F E Dwulet Journal: Transplant Proc Date: 1995-12 Impact factor: 1.066
Authors: A M James Shapiro; Camillo Ricordi; Bernhard J Hering; Hugh Auchincloss; Robert Lindblad; R Paul Robertson; Antonio Secchi; Mathias D Brendel; Thierry Berney; Daniel C Brennan; Enrico Cagliero; Rodolfo Alejandro; Edmond A Ryan; Barbara DiMercurio; Philippe Morel; Kenneth S Polonsky; Jo-Anna Reems; Reinhard G Bretzel; Federico Bertuzzi; Tatiana Froud; Raja Kandaswamy; David E R Sutherland; George Eisenbarth; Miriam Segal; Jutta Preiksaitis; Gregory S Korbutt; Franca B Barton; Lisa Viviano; Vicki Seyfert-Margolis; Jeffrey Bluestone; Jonathan R T Lakey Journal: N Engl J Med Date: 2006-09-28 Impact factor: 91.245
Authors: Marcio Fronza; Guilherme F Caetano; Marcel N Leite; Claudia S Bitencourt; Francisco W G Paula-Silva; Thiago A M Andrade; Marco A C Frade; Irmgard Merfort; Lúcia H Faccioli Journal: PLoS One Date: 2014-11-13 Impact factor: 3.240
Authors: Alisa M White; James G Shamul; Jiangsheng Xu; Samantha Stewart; Jonathan S Bromberg; Xiaoming He Journal: ACS Biomater Sci Eng Date: 2019-12-02
Authors: Robert W Holdcraft; Michael L Green; Andrew G Breite; Lisa Circle; Eric D Meyer; Hollie Adkins; Steven G Harbeck; Barry H Smith; Lawrence S Gazda Journal: Transplant Direct Date: 2016-06-06
Authors: Meirigeng Qi; Keiko Omori; Yoko Mullen; Brian McFadden; Luis Valiente; Jemily Juan; Shiela Bilbao; Bernard R Tegtmeier; Donald Dafoe; Fouad Kandeel; Ismail H Al-Abdullah Journal: Transplant Direct Date: 2016-02
Authors: Bashar Khiatah; Meirigeng Qi; Weiting Du; Kuan T-Chen; Kayleigh M van Megen; Rachel G Perez; Jeffrey S Isenberg; Fouad Kandeel; Bart O Roep; Hsun Teresa Ku; Ismail H Al-Abdullah Journal: Stem Cell Res Ther Date: 2019-11-15 Impact factor: 6.832
Authors: Nicole Corrales; Soomin Park; Hien Lau; Ivana Xu; Colleen Luong; Samuel Rodriguez; Johanna Mönch; Michael Alexander; Jonathan Rt Lakey Journal: Cell Transplant Date: 2020 Jan-Dec Impact factor: 4.064