Cryopreserved human fetal pancreas: a source of insulin-producing tissue?
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 degrees C, prior to stepwise digestion into 50- to 150-micron 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 degrees C, and a slow cooling rate of 0.25 degrees C/min to -40 degrees C, followed by submersion in liquid nitrogen (-196 degrees C). Rapid thawing at 300 degrees C/min from -196 degrees 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.
Dawidson
I
I
Department of Surgery, University of Texas Health Science Center, Dallas 75235.
Simonsen
R
R
Aggarwal
S
S
Coorpender
L
L
Diller
K
K
Rajotte
R
R
Raskin
P
P
Redman
H
H
Rosenstock
J
J
D000022:Abortion, Spontaneous; D000328:Adult; D002470:Cell Survival; D003922:Diabetes Mellitus, Type 1 / Q000628:therapy*; D004121:Dimethyl Sulfoxide; D005260:Female; D005615:Freezing; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000378:metabolism*; D008854:Microscopy, Electron; D010179:Pancreas / Q000166:cytology / Q000196:embryology; D016035:Pancreas Transplantation; D011247:Pregnancy; D014021:Tissue Preservation / Q000379:methods
D007328:Insulin; D004121:Dimethyl Sulfoxide