TY - JOUR
T1 - Cryopreserved human fetal pancreas
T2 - A source of insulin-producing tissue?
AU - Dawidson, Ingemar
AU - Simonsen, Randall
AU - Aggarwal, Shanti
AU - Coorpender, Laura
AU - Diller, Kenneth
AU - Rajotte, Ray
AU - Raskin, Philip
AU - Redman, Helen
AU - Rosenstock, Julio
N1 - Funding Information:
Received August 8, 1986; accepted September 14, 1987. ’ Supported by The University of Texas Health Science Center State Kidney and Pancreatic Unit, the Juvenile Diabetes Foundation, the Medical Research Council of Canada, the Muttart Diabetes Research and Training Center, and the National Diabetes Research Interchange. ‘To whom correspondence and reprint requests should be addressed at Department of Surgery, E7.202, University of Texas Health Science Center at Dallas, 5323 Harry Hines Blvd., Dallas, TX 75235.
PY - 1988/4
Y1 - 1988/4
N2 - Human fetal pancreata (HFP) were obtained from dilatation and extraction aborted fetuses of 11-18 weeks' gestation. The pancreas was excised under sterile conditions and kept in culture medium at 4 °C, prior to stepwise digestion into 50- to 150-μm fragments. The fragmented pieces were allowed to sediment by gravity, then transferred to tissue culture for 24-48 h, and cryopreserved. The freeze-thaw protocol used stepwise equilibration with dimethyl sulfoxide, nucleation of the sample at -10 °C, and a slow cooling rate of 0.25 °C/min to -40 °C, followed by submersion in liquid nitrogen (-196 °C). Rapid thawing at 300 °C/min from -196 °C was employed. Both fresh and frozen-thawed HFP fragments appeared viable as judged by light and electron microscopy, and secreted insulin in a perifusion system upon stimulation with glucose (28 mM) and theophylline (10 mM) or glucose (2.8 mM) and theophylline (10 mM). Six patients with Type I insulin-dependent diabetes mellitus, already requiring immunosuppression for a kidney transplant, had intraportal injection of 20 cryopreserved-thawed and pooled HFP fragments. Up to the 1-year post-transplant follow-up, there has been no evidence of in vivo insulin or C-peptide production. The usefulness of cryopreserved human fetal pancreata as a source of insulin-producing tissue for diabetic patients, therefore, remains to be demonstrated.
AB - Human fetal pancreata (HFP) were obtained from dilatation and extraction aborted fetuses of 11-18 weeks' gestation. The pancreas was excised under sterile conditions and kept in culture medium at 4 °C, prior to stepwise digestion into 50- to 150-μm fragments. The fragmented pieces were allowed to sediment by gravity, then transferred to tissue culture for 24-48 h, and cryopreserved. The freeze-thaw protocol used stepwise equilibration with dimethyl sulfoxide, nucleation of the sample at -10 °C, and a slow cooling rate of 0.25 °C/min to -40 °C, followed by submersion in liquid nitrogen (-196 °C). Rapid thawing at 300 °C/min from -196 °C was employed. Both fresh and frozen-thawed HFP fragments appeared viable as judged by light and electron microscopy, and secreted insulin in a perifusion system upon stimulation with glucose (28 mM) and theophylline (10 mM) or glucose (2.8 mM) and theophylline (10 mM). Six patients with Type I insulin-dependent diabetes mellitus, already requiring immunosuppression for a kidney transplant, had intraportal injection of 20 cryopreserved-thawed and pooled HFP fragments. Up to the 1-year post-transplant follow-up, there has been no evidence of in vivo insulin or C-peptide production. The usefulness of cryopreserved human fetal pancreata as a source of insulin-producing tissue for diabetic patients, therefore, remains to be demonstrated.
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U2 - 10.1016/0011-2240(88)90001-6
DO - 10.1016/0011-2240(88)90001-6
M3 - Article
C2 - 3286124
AN - SCOPUS:0023928869
SN - 0011-2240
VL - 25
SP - 83
EP - 93
JO - Cryobiology
JF - Cryobiology
IS - 2
ER -