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Purification of islets and cells from islets

Nielsen, J Höiriis and Lernmark, Åke LU orcid (1983) 2. p.99-126
Abstract
This chapter discusses the purification of islets and cells from islets. The pancreatic islets that are composed of several hormone-producing cells can be isolated by collagenase digestion of the pancreas. The yield of islets relative to the total number is usually small. Sufficient number of islets for a variety of functional and biochemical studies can be obtained by gradient centrifugations or by purification in organ culture. The isolated pancreatic islets are dispersed by enzymatic digestion or mechanical disruption of the islets in Ca2+free medium. The single cells are functionally intact and attempts are made to separate islet cell subpopulations by elutriation or gradient centrifugations, cell sorting, or other techniques such as... (More)
This chapter discusses the purification of islets and cells from islets. The pancreatic islets that are composed of several hormone-producing cells can be isolated by collagenase digestion of the pancreas. The yield of islets relative to the total number is usually small. Sufficient number of islets for a variety of functional and biochemical studies can be obtained by gradient centrifugations or by purification in organ culture. The isolated pancreatic islets are dispersed by enzymatic digestion or mechanical disruption of the islets in Ca2+free medium. The single cells are functionally intact and attempts are made to separate islet cell subpopulations by elutriation or gradient centrifugations, cell sorting, or other techniques such as free flow electrophoresis, sedimentation at unit gravity, or by immunocytochemical techniques. A major task in islet cell purification is to achieve preparations of highly purified endocrine cells and to separate the individual cell types from each other. Flow cytofluorophotometers, which can analyze and separate cells at a high speed based on either light scattering or fluorescence signals, have been found useful in attempts to separate islet cells. The fluorescence-activated cell sorter also allows analysis and sorting of islet cells based on indirect immunofluorescence. Tissue or cell-specific monoclonal antibodies represent useful reagents to allow sorting of certain cell types based on immunochemical labeling. A number of monoclonal antibodies reactive with specific cell-surface antigens on endocrine islet cells have been described in the chap
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Please use this url to cite or link to this publication:
author
and
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
host publication
Cell separation : methods and selected applications - methods and selected applications
editor
Pretlow, Thomas G and Pretlow, Theresa P
volume
2
pages
99 - 126
publisher
Academic Press
ISBN
978-0-12-564502-7
DOI
10.1016/B978-0-12-564502-7.50013-X
language
English
LU publication?
no
id
6e39df7d-10f8-4951-9fb3-8bcde95f6837
date added to LUP
2021-09-23 16:58:03
date last changed
2021-09-24 01:33:15
@inbook{6e39df7d-10f8-4951-9fb3-8bcde95f6837,
  abstract     = {{This chapter discusses the purification of islets and cells from islets. The pancreatic islets that are composed of several hormone-producing cells can be isolated by collagenase digestion of the pancreas. The yield of islets relative to the total number is usually small. Sufficient number of islets for a variety of functional and biochemical studies can be obtained by gradient centrifugations or by purification in organ culture. The isolated pancreatic islets are dispersed by enzymatic digestion or mechanical disruption of the islets in Ca2+free medium. The single cells are functionally intact and attempts are made to separate islet cell subpopulations by elutriation or gradient centrifugations, cell sorting, or other techniques such as free flow electrophoresis, sedimentation at unit gravity, or by immunocytochemical techniques. A major task in islet cell purification is to achieve preparations of highly purified endocrine cells and to separate the individual cell types from each other. Flow cytofluorophotometers, which can analyze and separate cells at a high speed based on either light scattering or fluorescence signals, have been found useful in attempts to separate islet cells. The fluorescence-activated cell sorter also allows analysis and sorting of islet cells based on indirect immunofluorescence. Tissue or cell-specific monoclonal antibodies represent useful reagents to allow sorting of certain cell types based on immunochemical labeling. A number of monoclonal antibodies reactive with specific cell-surface antigens on endocrine islet cells have been described in the chap<br/>}},
  author       = {{Nielsen, J Höiriis and Lernmark, Åke}},
  booktitle    = {{Cell separation : methods and selected applications}},
  editor       = {{Pretlow, Thomas G and Pretlow, Theresa P}},
  isbn         = {{978-0-12-564502-7}},
  language     = {{eng}},
  pages        = {{99--126}},
  publisher    = {{Academic Press}},
  title        = {{Purification of islets and cells from islets}},
  url          = {{http://dx.doi.org/10.1016/B978-0-12-564502-7.50013-X}},
  doi          = {{10.1016/B978-0-12-564502-7.50013-X}},
  volume       = {{2}},
  year         = {{1983}},
}