Development and functional characterization of extrahepatic cholangiocyte lines from normal rats.
BACKGROUND
Since limited in vitro tools exist for evaluating the pathophysiology of extrahepatic bile ducts, we aim to develop an extrahepatic cholangiocyte culture system from normal rats.


METHODS
Extrahepatic ducts were dissected from rats, cut in half length-wise and cultured on collagen-I coated plates. Transepithelial electrical resistance was measured. At ∼85% confluence, in extrahepatic cholangiocytes we measured: (i) cell size and distribution, and expression for cytokeratin-19, secretin, secretin receptor and somatostatin receptor type II (SSTR2), cystic fibrosis transmembrane conductance regulator (CFTR), chloride bicarbonate anion exchanger 2 (AE2), vascular endothelial growth factor-A (VEGF-A) and nerve growth factor (NGF); and (ii) the effect of secretin and/or somatostatin on 3'-5'-cyclic adenosine monophosphate (cAMP) levels and proliferation.


RESULTS
Cytokeratin-positive extrahepatic cholangiocytes were cultured for 6 passages to form a cell monolayer. Cholangiocytes proliferated to confluence over a 2-week period. The size of extrahepatic cholangiocytes averaged ∼16 μm. Extrahepatic ducts and cholangiocytes were positive for secretin, secretin receptor and SSTR2, CFTR, AE2, VEGF-A and NGF. In extrahepatic cholangiocyte cultures, secretin increased cAMP (prevented by somatostatin), chloride efflux and proliferation.


CONCLUSIONS
Extrahepatic cholangiocyte cultures may be important for studying diseases targeting extrahepatic cholangiocytes such as biliary atresia.
Department Medicine, Division Gastroenterology, S&W and Texas A&M HSC, Temple, TX, United States.
Julie
J
Venter
Research, Central Texas Veterans Health Care System, United States; Scott & White Digestive Disease Research Center, Academic Operations, Scott & White, United States; Department Medicine, Division Gastroenterology, S&W and Texas A&M HSC, Temple, TX, United States.
Heather
H
Francis
Research, Central Texas Veterans Health Care System, United States; Scott & White Digestive Disease Research Center, Academic Operations, Scott & White, United States; Department Medicine, Division Gastroenterology, S&W and Texas A&M HSC, Temple, TX, United States.
Fanyin
F
Meng
Research, Central Texas Veterans Health Care System, United States; Scott & White Digestive Disease Research Center, Academic Operations, Scott & White, United States; Department Medicine, Division Gastroenterology, S&W and Texas A&M HSC, Temple, TX, United States.
Sharon
S
DeMorrow
Research, Central Texas Veterans Health Care System, United States.
Lindsey
L
Kennedy
Research, Central Texas Veterans Health Care System, United States.
Holly
H
Standeford
Scott & White Digestive Disease Research Center, Academic Operations, Scott & White, United States.
Laura
L
Hargrove
Department Medicine, Division Gastroenterology, S&W and Texas A&M HSC, Temple, TX, United States.
Nan
N
Wu
Scott & White Digestive Disease Research Center, Academic Operations, Scott & White, United States.
Ying
Y
Wan
Department Medicine, Division Gastroenterology, S&W and Texas A&M HSC, Temple, TX, United States.
Gabriel
G
Frampton
Department Medicine, Division Gastroenterology, S&W and Texas A&M HSC, Temple, TX, United States.
Matthew
M
McMillin
Universita' Politecnica delle Marche, Ospedali Riuniti, Ancona, Italy.
Marco
M
Marzioni
Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, "Sapienza", Rome, Italy.
Eugenio
E
Gaudio
Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, "Sapienza", Rome, Italy.
Paolo
P
Onori
Research, Central Texas Veterans Health Care System, United States; Scott & White Digestive Disease Research Center, Academic Operations, Scott & White, United States; Department Medicine, Division Gastroenterology, S&W and Texas A&M HSC, Temple, TX, United States.
Shannon
S
Glaser
Research, Central Texas Veterans Health Care System, United States; Scott & White Digestive Disease Research Center, Academic Operations, Scott & White, United States; Department Medicine, Division Gastroenterology, S&W and Texas A&M HSC, Temple, TX, United States. Electronic address: galpini@tamu.edu.
Gianfranco
G
Alpini
D000818:Animals; D017734:Bile Ducts, Extrahepatic / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D018929:Cell Culture Techniques; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D027963:Chloride-Bicarbonate Antiporters / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism*; D017097:Electric Impedance; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D053539:Keratin-19 / Q000378:metabolism*; D020932:Nerve Growth Factor / Q000378:metabolism*; D051381:Rats; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D011964:Receptors, Gastrointestinal Hormone / Q000378:metabolism*; D017481:Receptors, Somatostatin / Q000378:metabolism*; D012633:Secretin / Q000378:metabolism* / Q000494:pharmacology; D013004:Somatostatin / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D027963:Chloride-Bicarbonate Antiporters; D053539:Keratin-19; D043562:Receptors, G-Protein-Coupled; D011964:Receptors, Gastrointestinal Hormone; D017481:Receptors, Somatostatin; D042461:Vascular Endothelial Growth Factor A; C029937:secretin receptor; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D012633:Secretin; D013004:Somatostatin; D020932:Nerve Growth Factor; C093733:somatostatin receptor 2; D000242:Cyclic AMP
Epithelia; Extrahepatic bile duct; Gastrointestinal hormones; Proliferation