BACKGROUND: The preservation of sufficient quantities of islets for human transplantation has proven to be a tenacious problem for researchers and transplant programs. Beyond the variables associated with islet procurement, there is the problem of tissue storage before transplantation. Cryopreservation has been adopted as a method for long-term islet storage that allows for recovery of viable tissue. However, there is significant tissue loss during the process and the possibility that long-term viability may be compromised. An alternate method of prolonged culture at 24 degrees C was initially introduced as a means of reducing islet antigenicity. Although successful in the short term, prolonged culture with serum-based media has also resulted in a significant loss of tissue. In this study, we report the successful use of an ITS+ Premix-supplemented serum-free media for prolonged islet culture and its comparison to fetal bovine serum-supplemented media and to cryopreservation. METHODS: Pancreata were procured from cadaveric organ donors, and islets were isolated using our own modification of the automated method of Ricordi. Aliquots from a series of human islet isolations were cultured in parallel in (A) CMRL + ITS (serum-free media; SFM) or (B) CMRL +10% fetal bovine serum (standard media) and compared with cryopreserved and thawed tissue. RESULTS: Our results show that SFM allows for the long-term culture of islet tissue. For time points up to 2 months, islets cultured in SFM showed recovery ratios greater than those for standard serum-supple. mented media. At 1 week and 1 month, islet recovery ratios were greater for SFM-cultured islets than for cryopreserved tissue. Viability studies confirmed that the SFM-cultured islets were able to respond to glucose stimulation (stimulation index 0.8-21.2). Additionally, in vivo results using cultured islets in a patient demonstrated good islet function, with a 1-month stimulation index of 4.02 in response to an intravenous glucose tolerance test. CONCLUSION: We conclude that this culture modification represents a method by which functional islet tissue can be maintained in long-term culture and successfully transplanted.
BACKGROUND: The preservation of sufficient quantities of islets for human transplantation has proven to be a tenacious problem for researchers and transplant programs. Beyond the variables associated with islet procurement, there is the problem of tissue storage before transplantation. Cryopreservation has been adopted as a method for long-term islet storage that allows for recovery of viable tissue. However, there is significant tissue loss during the process and the possibility that long-term viability may be compromised. An alternate method of prolonged culture at 24 degrees C was initially introduced as a means of reducing islet antigenicity. Although successful in the short term, prolonged culture with serum-based media has also resulted in a significant loss of tissue. In this study, we report the successful use of an ITS+ Premix-supplemented serum-free media for prolonged islet culture and its comparison to fetal bovine serum-supplemented media and to cryopreservation. METHODS: Pancreata were procured from cadaveric organ donors, and islets were isolated using our own modification of the automated method of Ricordi. Aliquots from a series of human islet isolations were cultured in parallel in (A) CMRL + ITS (serum-free media; SFM) or (B) CMRL +10% fetal bovine serum (standard media) and compared with cryopreserved and thawed tissue. RESULTS: Our results show that SFM allows for the long-term culture of islet tissue. For time points up to 2 months, islets cultured in SFM showed recovery ratios greater than those for standard serum-supple. mented media. At 1 week and 1 month, islet recovery ratios were greater for SFM-cultured islets than for cryopreserved tissue. Viability studies confirmed that the SFM-cultured islets were able to respond to glucose stimulation (stimulation index 0.8-21.2). Additionally, in vivo results using cultured islets in a patient demonstrated good islet function, with a 1-month stimulation index of 4.02 in response to an intravenous glucose tolerance test. CONCLUSION: We conclude that this culture modification represents a method by which functional islet tissue can be maintained in long-term culture and successfully transplanted.
Authors: Priya M Miranda; Viswanathan Mohan; Sekhar Ganthimathy; Ranjit M Anjana; S Gunasekaran; Venkatachalam Thiagarajan; Thomas A Churchill; Tatsuya Kin; A M James Shapiro; Jonathan R T Lakey Journal: Islets Date: 2013 Sep-Dec Impact factor: 2.694
Authors: Gopalakrishnan Loganathan; Rajinder K Dawra; Subbiah Pugazhenthi; Zhiguang Guo; Sajjad M Soltani; Alexander Wiseman; Mark A Sanders; Klearchos K Papas; Kumaravel Velayutham; Ashok K Saluja; David E R Sutherland; Bernhard J Hering; A N Balamurugan Journal: Transplantation Date: 2011-12-15 Impact factor: 4.939
Authors: Yi-Ju Chen; Taiji Yamazoe; Karla F Leavens; Fabian L Cardenas-Diaz; Andrei Georgescu; Dongeun Huh; Paul Gadue; Ben Z Stanger Journal: JCI Insight Date: 2019-11-01
Authors: Ajit S Narang; Kun Cheng; James Henry; Chunxiang Zhang; Omaima Sabek; Daniel Fraga; Malak Kotb; A Osama Gaber; Ram I Mahato Journal: Pharm Res Date: 2004-01 Impact factor: 4.200
Authors: Jocelyn E Manning Fox; James Lyon; Xiao Qing Dai; Robert C Wright; Julie Hayward; Martijn van de Bunt; Tatsuya Kin; A M James Shapiro; Mark I McCarthy; Anna L Gloyn; Mark D Ungrin; Jonathan R Lakey; Norm M Kneteman; Garth L Warnock; Gregory S Korbutt; Raymond V Rajotte; Patrick E MacDonald Journal: Diabetologia Date: 2015-05-01 Impact factor: 10.122