Cryopreserved human fetal pancreas: A source of insulin-producing tissue?

Ingemar Dawidson, Randall Simonsen, Shanti Aggarwal, Laura Coorpender, Kenneth Diller, Ray Rajotte, Philip Raskin, Helen Redman, Julio Rosenstock

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4 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)83-93
Number of pages11
JournalCryobiology
Volume25
Issue number2
DOIs
StatePublished - Apr 1988

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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    Dawidson, I., Simonsen, R., Aggarwal, S., Coorpender, L., Diller, K., Rajotte, R., Raskin, P., Redman, H., & Rosenstock, J. (1988). Cryopreserved human fetal pancreas: A source of insulin-producing tissue? Cryobiology, 25(2), 83-93. https://doi.org/10.1016/0011-2240(88)90001-6