Joersbo, M, and Brunstedt, J, 1992, Sonication: A new method for gene transfer to plants. -Physiol. Plant. 85: 230-234.Sonieation is a novel method for gene transfer into plant protoplasts and intact plant cells. The mode of action of ultrasound and its chemical, biochemical and physiological effects, are reviewed. The state of the art of acotistic transformation is presented and possible mechanisms are discussed.
Islets of Langerhans were isolated from four human kidney donors, aged 16 to 21 years by the collagenase method described for isolation of rodent islets. So far the human islets have been kept in tissue culture, without attachment, in medium RPMI 1640 supplemented with 10% calf serum for more than 9 months, with preservation of the ability to release insulin in response to glucose stimulation. Replacement of calf serum with serum from normal human subjects did not affect B-cell survival, but resulted in elevated insulin values partly due to lower insulin degrading activity. Thus the described technique presents a valuable tool for studying chronic effects of metabolites and hormones on islet function, as well as for islet storage prior to transplantation into humans.
Abstract.
The effects of glucocorticoids on the pancreatic endocrine function was studied in isolated mouse pancreatic islets maintained in tissue culture for 1 to 3 weeks. Following culture for 1 week without corticoid supplement acute experiments with hydrocortisone showed no significant effect on the glucose-induced insulin release at 10−8 to 10−5 mol/l hydrocortisone. When, however, the islets were cultured in the presence of hvdrocortisone, there was an increased insulin release to the medium in a dose-dependent manner, with the maximal effect at 10−7 mol/l hydrocortisone. The release of glucagon to the medium was not affected to the same degree, but showed a slight inhibition at increasing concentrations of hydrocortisone.
Short-term experiments after the culture period showed that islets cultured for 3 weeks in the presence of 10−7 to 10−5 mol/l hydrocortisone had an enhanced insulin secretion in response to glucose. The islets did not show any statistically significant change in their insulin- and DNA-content after 3 weeks of culture with hydrocortisone, but a marked reduction in the content of glucagon was found with increasing concentrations of hydrocortisone.
The present results suggest that physiological concentrations of hydrocortisone are of importance for mouse islets to maintain their insulin production in tissue culture.
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