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The preparation of, and studies on, free cell suspensions from mouse pancreatic islets

Lernmark, Å LU orcid (1974) In Diabetologia 10(5). p.431-438
Abstract

Pancreatic islets, isolated from the pancreas of obese-hyperglycemic mice, were used to prepare free islet cells in suspension. Batches of 100 islets were disrupted by mechanical shaking for 10 sec in a Ca2+-free HEPES-buffered Krebs-Ring'er medium containing 1 mM EGTA. From 200-500 islets, about 2.2×106 cells could be obtained in suspension, corresponding to a yield of roughly 55% as calculated from the content of DNA in cells and islets. The isolated islet cells appeared well-preserved in the light and the electron microscope and seemed to exhibit a high degree of viability as judged by viable cell counts, a radioactive assay for lysis and insulin release. Insulin release from islet cell suspensions, as... (More)

Pancreatic islets, isolated from the pancreas of obese-hyperglycemic mice, were used to prepare free islet cells in suspension. Batches of 100 islets were disrupted by mechanical shaking for 10 sec in a Ca2+-free HEPES-buffered Krebs-Ring'er medium containing 1 mM EGTA. From 200-500 islets, about 2.2×106 cells could be obtained in suspension, corresponding to a yield of roughly 55% as calculated from the content of DNA in cells and islets. The isolated islet cells appeared well-preserved in the light and the electron microscope and seemed to exhibit a high degree of viability as judged by viable cell counts, a radioactive assay for lysis and insulin release. Insulin release from islet cell suspensions, as calculated per cell, per content of DNA or insulin or per packed cell volume, was stimulated by glucose alone or in combination with theophylline. The glucose response was low compared with that of intact isolated islets.

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Please use this url to cite or link to this publication:
author
publishing date
type
Contribution to journal
publication status
published
keywords
DNA content, insulin release, islet disruption, Isolated islet cells, ob/ob mouse islets
in
Diabetologia
volume
10
issue
5
pages
431 - 438
publisher
Springer
external identifiers
  • pmid:4375640
  • scopus:0016209124
ISSN
0012-186X
DOI
10.1007/BF01221634
language
English
LU publication?
no
id
ebd559c3-d0cf-4ffa-8a2f-465eadf4fe8a
date added to LUP
2019-09-18 12:17:03
date last changed
2024-03-13 08:10:15
@article{ebd559c3-d0cf-4ffa-8a2f-465eadf4fe8a,
  abstract     = {{<p>Pancreatic islets, isolated from the pancreas of obese-hyperglycemic mice, were used to prepare free islet cells in suspension. Batches of 100 islets were disrupted by mechanical shaking for 10 sec in a Ca<sup>2+</sup>-free HEPES-buffered Krebs-Ring'er medium containing 1 mM EGTA. From 200-500 islets, about 2.2×10<sup>6</sup> cells could be obtained in suspension, corresponding to a yield of roughly 55% as calculated from the content of DNA in cells and islets. The isolated islet cells appeared well-preserved in the light and the electron microscope and seemed to exhibit a high degree of viability as judged by viable cell counts, a radioactive assay for lysis and insulin release. Insulin release from islet cell suspensions, as calculated per cell, per content of DNA or insulin or per packed cell volume, was stimulated by glucose alone or in combination with theophylline. The glucose response was low compared with that of intact isolated islets.</p>}},
  author       = {{Lernmark, Å}},
  issn         = {{0012-186X}},
  keywords     = {{DNA content; insulin release; islet disruption; Isolated islet cells; ob/ob mouse islets}},
  language     = {{eng}},
  month        = {{10}},
  number       = {{5}},
  pages        = {{431--438}},
  publisher    = {{Springer}},
  series       = {{Diabetologia}},
  title        = {{The preparation of, and studies on, free cell suspensions from mouse pancreatic islets}},
  url          = {{http://dx.doi.org/10.1007/BF01221634}},
  doi          = {{10.1007/BF01221634}},
  volume       = {{10}},
  year         = {{1974}},
}