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A lack of a sufficient number of human donor pancreases has stimulated interest in isolation and cryopreservation techniques for islets from the porcine pancreas. But because of a poorly developed outer membrane porcine islets are particularly susceptible to damage during cryopreservation. The aims of this study were two-fold: 1) to develop a method for isolation and storage of islets from neonatal porcine pancreas and, 2) to examine effects of Sertoli cells on islet yield and function in Sertoli cell-islet cell cocultures. A total of 170 neonatal porcine pancreases were processed by means of a short period of digestion with collagenase and culture of the tissues at 32 degrees C for periods up to 7 days following isolation. Results were: The mean +/- SEM, number of viable islets, and percentage loss of cells following 7 days of culture were 29,442 +/- 1,119 and 22.2 +/- 1.2, respectively, Cryopreservation had a marked impact on recovery of viable islets: In absence of Sertoli cells an average of only 64% of islets remained viable; by contrast, when cryopreserved islets were cocultured with Sertoli cells, a mean of 82% was recovered. Glucose at 16.7 mmol/L had the capacity to elicit insulin release from 3-day-old cultured islets. The concentration in absence of Sertoli cells was 57.3 +/- 3.8 uU/mL/10 islets; in the presence of Sertoli cells the level increased to a mean +/- SEM of 112.8 +/- 17.7, uU/mL/10 islets. Similar results were obtained following cryopreservation: glucose at 16.7 mmol/L stimulated a mean +/- SEM of 27.9 +/- 6.6, uU/mL/10 islets, of insulin in absence of, and 44.9 +/- 9.9, uU/mL/10 islets, in presence of, Sertoli cells. Our results show that isolation and cryopreservation of neonatal porcine islets can be successfully accomplished. In addition, coculture with Sertoli cells significantly improves both the yield and functional capacity of islets following cryopreservation.

Hematopoietic cell transplantation (HCT) confers a longterm disease-modifying therapy for transplant-permissive inherited metabolic diseases (IMD). We examined the outcome of children with IMD received first HCT at Royal Manchester ChildrenÕs hospital from 1985 to 2016, a national metabolic transplant center in the United Kingdom. Cox proportional-hazard models were used to examine the predictors for overall survival. 137 children with IMD were included in this analysis (historical cohort (1985-2006), n = 65; current cohort (2007)(2008)(2009)(2010)(2011)(2012)(2013)(2014)(2015)(2016), n = 72). Primary diagnoses were mucopolysaccharidoses (MPS, n = 112, 81%; MPS IH, n = 103, MPS II, n = 1, MPS III = 2, MPS VI = 6), X-linked adrenoleukodystrophy (XALD, n = 8, 6%), metachromatic leukodystrophy (n = 5, 4%), alpha mannosidosis (n = 4, 3%), Wolman disease (n = 3, 2%) and others (n = 5, 4%). The median age at transplant was 14.4 (range 1.6-220.7) months. Fifty-four children (40%) received relative donor HCT (marrow, n = 43 (78%), apheresis, n = 11 (20%) and cord blood (CB) = 2 (2%)) while 83 (60%) had unrelated donor HCT (marrow, n = 31 (37%), apheresis, n = 9 (11%) and CB, n = 43 (52%)). The estimated 5-year OS improved from 65% (95% CI, 53-76%) in the historical cohort to 91% (95%CI, 81-96%) in the current cohort (P < .001). Male (HR .35, 95%CI, .15-.81, P = .01) and grade III-IV GVHD (HR 5.10, 95%CI, 1.72-15.14, P = .003) were predictors for overall survival. In current cohort, none of the pre-transplant factors, transplant factors and transplant related complications were associated with OS. The estimated 5-year OS in children transplanted in the current cohort was 100% for XALD (n = 5), 98% (95% CI, 83%-98%) for MPS IH (n = 55) and 75% (95% CI, 42-92%) for other IMD. The estimated one-year TRM reduced from 23% (95% CI, 13-40%) in the historical cohort to 7% (95% CI, 3-18%) for current cohort (P = .006). The estimated 5-year engrafted survival (ES) has doubled from 41% (95% CI, 29-53%) in the historical cohort to 86% (95% CI, 75-92%) in the current cohort (P < .001). The proportion of patients with graft failure has decreased from 37% in the historical cohort to 8% in the current cohort (P < .001). Thirteen out of 18 patients (71%) in the historical cohort and all four patients (100%) in the current cohort receiving a second transplant were alive-and-engrafted. Of 82 survivors followed up at Manchester, 80% had full chimerism (_95% donor chimerism) while 20% had mixed chimerism (20-94% donor chimerism). Despite the limitation of a single centre, our findings reiterated the observation of HCT is a safe and life-long therapy for children with IMD. The outcome of a second transplant is good. S396Abstracts / Biol Blood Marrow Transplant 24 (2018) S291-S459 Conclusion: TNC is better than CD34 + cell doses for predicting transplantation outcomes after PBSC HCT in children. Further studies are indicated to validate these results.

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