title: Long-term cultured neonatal islet cell clusters demonstrate better outcomes for reversal of diabetes: in vivo and molecular profiles.
abstract: BACKGROUND
  Porcine neonatal islet-like cell clusters (NICC) are being considered as a source of β-cell replacement. However, the lag time to full function due to hormonal immaturity remains a problem. This study aimed to determine whether time in culture was important for NICC function in vivo.
  
  
  METHODS
  Neonatal islet-like cell clusters were isolated from piglets aged between 1 and 3 days, and cultured for up to 27 days post-isolation. Each week, NICC number, viability, and function were determined.
  
  
  RESULTS
  Neonatal islet-like cell clusters cultured for 12, 19, and 27 days achieved normal blood glucose levels at 46 days (85% of animals), 32 days (100% of animals), and 35 days (81% of animals), respectively. By comparison, standard 6-day culture took a mean of 63 days to achieve normoglycemia in 35% of animals. Longer time in culture resulted in a significant loss of islet equivalent over time. However, insulin gene expression levels were significantly higher at days 12, 19, 27 compared to day 6. Glucagon gene expression was highest at day 12, and significantly higher than day 6 at all time points. Bcl-2 gene expression increased over time, and tissue factor (TF) gene expression was highest on day 6 and then decreased over the remaining time points.
  
  
  CONCLUSIONS
  Culture of NICC for 12 days provides the best balance in vivo functional outcome for transplantation, shown by better reversal of diabetes, and higher levels of gene expression for insulin, glucagon and Bcl-2 and lower levels of TF expression with acceptable NICC number loss in terms of time and expense.
authors: The Centre for Transplant & Renal Research, Westmead Millennium Research Institute, University of Sydney, Westmead, NSW, Australia.
authors: Elvira
authors: E
authors: Jimenez-Vera
authors: Sussan
authors: S
authors: Davies
authors: Peta
authors: P
authors: Phillips
authors: Philip J
authors: PJ
authors: O'Connell
authors: Wayne J
authors: WJ
authors: Hawthorne
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001786:Blood Glucose / Q000378:metabolism; D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000601:surgery*; D005260:Female; D015870:Gene Expression; D019254:Genes, bcl-2; D005934:Glucagon / Q000235:genetics; D064593:Heterografts; D007328:Insulin / Q000235:genetics; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D016381:Islets of Langerhans Transplantation / Q000379:methods*; D008297:Male; D051379:Mice; D016513:Mice, SCID; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034421:Sus scrofa; D013925:Thromboplastin / Q000235:genetics; D013997:Time Factors
chemical_list: D001786:Blood Glucose; D007328:Insulin; D012333:RNA, Messenger; D005934:Glucagon; D013925:Thromboplastin
keywords: islet cell culture; molecular profiling; neonatal islet cell clusters; type 1 diabetes; xenotransplantation