Improved method for isolation of porcine neonatal pancreatic cell clusters

Abstract

The technique allowed us to obtain NPCC with optimal viability, functionality, purity, and endurance for use in research.

Long-term follow-up of patients with type 1 diabetes transplanted with neonatal pig islets

“…[29] reported survival of 9·5 years of porcine islets xenotransplanted in one patient, who reduced his insulin dose by 30% but returned to pretransplant level at week 49; urinary porcine C‐peptide was detectable in the first 11 months at similar levels to those found in our patients post‐stimulation. Our results suggest prolonged survival of the implanted cells using the device [30,31] by the presence of urinary porcine C‐peptide, the important insulin dose reduction, the immunological response and biopsies positive for porcine islets, as well as initial excellent metabolic control. In basal conditions our patients produced small quantities of porcine C‐peptide, but this increased after stimulation with l ‐arginine, although even this increase is below normal levels of human urinary C‐peptide (65–262 µg/day).…”

Real‐time assessment of encapsulated neonatal porcine islets prior to clinical xenotransplantation

“…Seven-to-ten-day-old commercial newborn piglets (Pig Improvement Company, Maramarua, New Zealand) were anaesthetized, exsanguinated, and pancreata were removed using sterile techniques. Pancreata were processed as previously described [17]. Briefly, neonatal porcine islets were isolated using standard techniques of collagenase (Liberase MTF C/T, Roche, GMP grade) digestion and cultured in RPMI-1640 medium.…”

Size‐related assessment on viability and insulin secretion of caprine islets in vitro

“…We investigated the effect of caprine islet size on function of the islets in vitro culture 48 h after isolation. Small‐sized caprine islets consistently released an approximately 3‐fold higher amount of insulin than large‐sized islets under low and high glucose conditions during static incubation, which is an effective functional indicator for isolated islets . Previous findings by Huang et al.…”

“…We investigated the effect of caprine islet size on function of the islets in vitro culture 48 h after isolation. Small-sized caprine islets consistently released an approximately 3-fold higher amount of insulin than large-sized islets under low and high glucose conditions during static incubation, which is an effective functional indicator for isolated islets [22][23][24]. Previous findings by Huang et al [15] that small-sized islets had a higher density of cells/area than large-sized islets in vitro could be one of the plausible explanations for the higher insulin secretion from smallsized islets [15].…”