Human islet purification: a prospective comparison of Euro-Ficoll and bovine serum albumin density gradients.
Euro-Ficoll (EF) and bovine serum albumin (BSA) are the two most commonly used media for the density gradient purification of human pancreatic islets. The aim of this study was to compare these two media with respect to the efficiency of human islet isolation. Ten human pancreata were collagenase-digested, and samples of digest were separated on either a continuous linear density gradient of BSA or a discontinuous gradient of EF (1.108/1.096/1.037/Euro-Collins). Efficiency of islet purification was assessed by insulin and amylase assay of aliquots aspirated from the BSA gradients, and from the interfaces of the EF gradients. Islets were obtained from two interfaces in the EF gradients. Islet yield from the upper interface was generally poor (median 28% of total insulin; range 2-71%), but purity was better than for an equivalent yield using BSA [1% (0-3%) amylase contamination for EF versus 6% (0-37%) for BSA; P = 0.013]. Pooling both EF interfaces increased yield to 66% (17-81%) but markedly reduced purity [46% (0-50%) amylase for EF versus 31% (0-52%) for BSA]. In conclusion, the efficiency of human islet purification is similar, though disappointingly low, with BSA and with EF. Considerable scope exists, therefore, for improvement in the density gradient purification of human islets.
Chadwick
D R
DR
Department of Surgery, Leicester Royal Infirmary, UK.
Robertson
G S
GS
Contractor
H
H
Swift
S
S
Rose
S
S
Thirdborough
S T
ST
Chamberlain
R
R
James
R F
RF
Bell
P R
PR
London
N J
NJ
D000293:Adolescent; D000328:Adult; D001926:Brain Death; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000379:methods; D005362:Ficoll; D006801:Humans; D006982:Hypertonic Solutions; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D008875:Middle Aged; D012710:Serum Albumin, Bovine; D014019:Tissue Donors
D006982:Hypertonic Solutions; D007328:Insulin; D005362:Ficoll; D012710:Serum Albumin, Bovine