Duct Epithelial Cells Cultured from Human Pancreas Processed for Transplantation Retain Differentiated Ductal Characteristics

Carol Kolar, Thomas Caffrey, Michael Hollingsworth, Mark Scheetz, Marie Sutherlin, Lamont Weide, Terence Lawson

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Summary: A procedure is described for the isolation and growth in vitro of epithelial cells from the duct network of human pancreas, referred to as DEC. A significant advantage of our procedure over previously published procedures is that it enables the isolation of DEC from small pieces of pancreas tissue (<5 g) and, also, from the digest remaining after the isolation of islet cells from human pancreas, material that would normally be discarded. These were the only reliable sources for pancreas tissue available to us. This procedure shows that some of the techniques that have been successfully used for the isolation of rodent DEC are also valuable in the isolation of human DEC. In particular, the use of cholera toxin to prevent fibroblast growth and contamination obviates the need for the time-consuming procedure of physically removing fibroblasts or the use of expensive fibroblast-specific monoclo-nal antibodies. The use of sieving to separate the digest immediately achieves a partial purification, which, coupled with that of allowing duct cysts to form, adds to the purity of the final preparation. The ductal system of the intact pancreas tissue and the DEC derived from it expressed cytokeratins 7, 8/18, and 19 and markers for the presence of MUC1, CFTR, and carbonic anhydrase II, which are specific for ductal epithelial cells or for pancreatic ductal functions. This study showed that it is possible to obtain selectively viable DEC from small ducts in otherwise waste pieces of human pancreas. It showed that these cells retained all of the epithelial characteristics that were examined and, in combination with data from an earlier study, showed that the cultured DEC retain the metabolic functions of duct epithelial cells in vivo.

Original languageEnglish (US)
Pages (from-to)265-271
Number of pages7
JournalPancreas
Volume15
Issue number3
DOIs
StatePublished - Jan 1 1997

Fingerprint

Pancreas Transplantation
Pancreas
Epithelial Cells
Fibroblasts
Carbonic Anhydrase II
Keratin-8
Keratin-7
Cholera Toxin
Growth
Islets of Langerhans
Cysts
Rodentia
Antibodies

Keywords

  • CFTR
  • Cytokeratin
  • Human
  • Pancreas duct epithelium
  • Tissue culture

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism
  • Hepatology
  • Endocrinology

Cite this

Duct Epithelial Cells Cultured from Human Pancreas Processed for Transplantation Retain Differentiated Ductal Characteristics. / Kolar, Carol; Caffrey, Thomas; Hollingsworth, Michael; Scheetz, Mark; Sutherlin, Marie; Weide, Lamont; Lawson, Terence.

In: Pancreas, Vol. 15, No. 3, 01.01.1997, p. 265-271.

Research output: Contribution to journalArticle

Kolar, Carol ; Caffrey, Thomas ; Hollingsworth, Michael ; Scheetz, Mark ; Sutherlin, Marie ; Weide, Lamont ; Lawson, Terence. / Duct Epithelial Cells Cultured from Human Pancreas Processed for Transplantation Retain Differentiated Ductal Characteristics. In: Pancreas. 1997 ; Vol. 15, No. 3. pp. 265-271.
@article{73bcc9916ac74ac18340a06b33f366d4,
title = "Duct Epithelial Cells Cultured from Human Pancreas Processed for Transplantation Retain Differentiated Ductal Characteristics",
abstract = "Summary: A procedure is described for the isolation and growth in vitro of epithelial cells from the duct network of human pancreas, referred to as DEC. A significant advantage of our procedure over previously published procedures is that it enables the isolation of DEC from small pieces of pancreas tissue (<5 g) and, also, from the digest remaining after the isolation of islet cells from human pancreas, material that would normally be discarded. These were the only reliable sources for pancreas tissue available to us. This procedure shows that some of the techniques that have been successfully used for the isolation of rodent DEC are also valuable in the isolation of human DEC. In particular, the use of cholera toxin to prevent fibroblast growth and contamination obviates the need for the time-consuming procedure of physically removing fibroblasts or the use of expensive fibroblast-specific monoclo-nal antibodies. The use of sieving to separate the digest immediately achieves a partial purification, which, coupled with that of allowing duct cysts to form, adds to the purity of the final preparation. The ductal system of the intact pancreas tissue and the DEC derived from it expressed cytokeratins 7, 8/18, and 19 and markers for the presence of MUC1, CFTR, and carbonic anhydrase II, which are specific for ductal epithelial cells or for pancreatic ductal functions. This study showed that it is possible to obtain selectively viable DEC from small ducts in otherwise waste pieces of human pancreas. It showed that these cells retained all of the epithelial characteristics that were examined and, in combination with data from an earlier study, showed that the cultured DEC retain the metabolic functions of duct epithelial cells in vivo.",
keywords = "CFTR, Cytokeratin, Human, Pancreas duct epithelium, Tissue culture",
author = "Carol Kolar and Thomas Caffrey and Michael Hollingsworth and Mark Scheetz and Marie Sutherlin and Lamont Weide and Terence Lawson",
year = "1997",
month = "1",
day = "1",
doi = "10.1097/00006676-199710000-00008",
language = "English (US)",
volume = "15",
pages = "265--271",
journal = "Pancreas",
issn = "0885-3177",
publisher = "Lippincott Williams and Wilkins",
number = "3",

}

TY - JOUR

T1 - Duct Epithelial Cells Cultured from Human Pancreas Processed for Transplantation Retain Differentiated Ductal Characteristics

AU - Kolar, Carol

AU - Caffrey, Thomas

AU - Hollingsworth, Michael

AU - Scheetz, Mark

AU - Sutherlin, Marie

AU - Weide, Lamont

AU - Lawson, Terence

PY - 1997/1/1

Y1 - 1997/1/1

N2 - Summary: A procedure is described for the isolation and growth in vitro of epithelial cells from the duct network of human pancreas, referred to as DEC. A significant advantage of our procedure over previously published procedures is that it enables the isolation of DEC from small pieces of pancreas tissue (<5 g) and, also, from the digest remaining after the isolation of islet cells from human pancreas, material that would normally be discarded. These were the only reliable sources for pancreas tissue available to us. This procedure shows that some of the techniques that have been successfully used for the isolation of rodent DEC are also valuable in the isolation of human DEC. In particular, the use of cholera toxin to prevent fibroblast growth and contamination obviates the need for the time-consuming procedure of physically removing fibroblasts or the use of expensive fibroblast-specific monoclo-nal antibodies. The use of sieving to separate the digest immediately achieves a partial purification, which, coupled with that of allowing duct cysts to form, adds to the purity of the final preparation. The ductal system of the intact pancreas tissue and the DEC derived from it expressed cytokeratins 7, 8/18, and 19 and markers for the presence of MUC1, CFTR, and carbonic anhydrase II, which are specific for ductal epithelial cells or for pancreatic ductal functions. This study showed that it is possible to obtain selectively viable DEC from small ducts in otherwise waste pieces of human pancreas. It showed that these cells retained all of the epithelial characteristics that were examined and, in combination with data from an earlier study, showed that the cultured DEC retain the metabolic functions of duct epithelial cells in vivo.

AB - Summary: A procedure is described for the isolation and growth in vitro of epithelial cells from the duct network of human pancreas, referred to as DEC. A significant advantage of our procedure over previously published procedures is that it enables the isolation of DEC from small pieces of pancreas tissue (<5 g) and, also, from the digest remaining after the isolation of islet cells from human pancreas, material that would normally be discarded. These were the only reliable sources for pancreas tissue available to us. This procedure shows that some of the techniques that have been successfully used for the isolation of rodent DEC are also valuable in the isolation of human DEC. In particular, the use of cholera toxin to prevent fibroblast growth and contamination obviates the need for the time-consuming procedure of physically removing fibroblasts or the use of expensive fibroblast-specific monoclo-nal antibodies. The use of sieving to separate the digest immediately achieves a partial purification, which, coupled with that of allowing duct cysts to form, adds to the purity of the final preparation. The ductal system of the intact pancreas tissue and the DEC derived from it expressed cytokeratins 7, 8/18, and 19 and markers for the presence of MUC1, CFTR, and carbonic anhydrase II, which are specific for ductal epithelial cells or for pancreatic ductal functions. This study showed that it is possible to obtain selectively viable DEC from small ducts in otherwise waste pieces of human pancreas. It showed that these cells retained all of the epithelial characteristics that were examined and, in combination with data from an earlier study, showed that the cultured DEC retain the metabolic functions of duct epithelial cells in vivo.

KW - CFTR

KW - Cytokeratin

KW - Human

KW - Pancreas duct epithelium

KW - Tissue culture

UR - http://www.scopus.com/inward/record.url?scp=0031252889&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031252889&partnerID=8YFLogxK

U2 - 10.1097/00006676-199710000-00008

DO - 10.1097/00006676-199710000-00008

M3 - Article

C2 - 9336790

AN - SCOPUS:0031252889

VL - 15

SP - 265

EP - 271

JO - Pancreas

JF - Pancreas

SN - 0885-3177

IS - 3

ER -