Effect of human islet rescue gradient purification on islet yield and fractional Beta cell viability.
It is difficult to consistently obtain sufficient postpurification islet numbers from younger donors because of the higher proportion of trapped islets after pancreas digestion. Continuous gradient purification (CGP), which is currently used in several islet processing centers, sometimes is not efficient in the purification of trapped islets. Rescue gradient purification (RGP) could improve postpurification islet yields, resulting in an increased number of islet cell products that could be transplanted. Sixty-eight human islet isolations, in which CGP was followed by RGP were analyzed. The quality of islets from CGP and RGP was assessed by beta-cell fractional viability (betaFV) and ADP/ATP ratio. Donor age negatively correlated with the proportion of the islets rescued by RGP (R = -0.52; P < .01) and to the percentage of trapped islets (R = -0.46; P < .01). Age-related differences were observed in pancreas weight, digestion time, and islet yields from CGP, respectively. Importantly, islets from RGP had an 11% higher betaFV compared with islets from CGP. ADP/ATP ratio was also lower in RGP islets versus CGP islets. RGP improved the efficiency of islet purification from younger pancreata and did not affect islet cell viability, which was actually higher in RGP fractions, indicating that rescued trapped islets from the pellet and lower purity layers are not damaged by the extra purification step and may actually be more viable. RGP could be used to rescue high-quality islets from less than 30% pure islet fractions, which are often discarded in the clinical setting.
Miki
A
A
Cell Transplant Center, Diabetes Research Institute, University of Miami Leonard M. Miller School of Medicine, Miami, Florida 33136, USA.
Ricordi
C
C
Yamamoto
T
T
Mita
A
A
Barker
S
S
Khan
A
A
Alejandro
R
R
Ichii
H
H
D000244:Adenosine Diphosphate / Q000032:analysis; D000255:Adenosine Triphosphate / Q000032:analysis; D000367:Age Factors; D002102:Cadaver; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology*; D007515:Islets of Langerhans / Q000166:cytology*; D014019:Tissue Donors
D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate