Establishment of a novel collagenase perfusion method to isolate rat pancreatic stellate cells and investigation of their gene expression of TGF-beta1, type I collagen, and CTGF in primary culture or freshly isolated cells.
In studies of the pathogenesis of pancreatic fibrosis, pancreatic stellate cells (PSCs) have recently gained attention. In the present study, we established a new collagenase perfusion method through thoracic aorta cannulation to isolate PSCs, and we studied gene expression of TGF-beta1, type I collagen, and connective tissue growth factor using primary cultured PSCs. Our method facilitated PSC isolation, and by our new method, 4.3 +/- 1.2 x 10(6) PSCs were obtained from a rat. In comparing the expression of these genes with that of hepatic stellate cells (HSCs), we observed a similar pattern, although PSCs expressed type I collagen gene earlier than did HSCs. These results suggest that PSCs may play an important role in fibrosis of the pancreas, as HSCs do in liver fibrosis; in addition, PSCs may exist in a preactivated state or may be more easily activated than are HSCs. We also isolated the PSCs from a WBN/Kob rat, the spontaneous pancreatitis rat, and compared the gene expression with that from a normal rat.
Shinji
Toshiyuki
T
Department of Laboratory Medicine, Okayama University Graduate School of Medicine and Dentistry, Japan. toshinji@md.okayama-u.ac.jp
Ujike
Kozo
K
Ochi
Koji
K
Kusano
Nobuchika
N
Kikui
Tetsuya
T
Matsumura
Naoki
N
Emori
Yasuyuki
Y
Seno
Toshinobu
T
Koide
Norio
N
D000818:Animals; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics*; D017364:Collagenases; D055513:Connective Tissue Growth Factor; D005355:Fibrosis; D015870:Gene Expression; D017874:Immediate-Early Proteins / Q000235:genetics*; D007158:Immunologic Techniques; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics*; D008297:Male; D008858:Microscopy, Phase-Contrast; D010179:Pancreas / Q000166:cytology* / Q000473:pathology / Q000502:physiology*; D010477:Perfusion / Q000379:methods*; D051381:Rats; D011919:Rats, Inbred Strains; D017208:Rats, Wistar; D016212:Transforming Growth Factor beta / Q000235:genetics*; D053773:Transforming Growth Factor beta1
C526316:CCN2 protein, rat; D024042:Collagen Type I; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; C508962:Tgfb1 protein, rat; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D055513:Connective Tissue Growth Factor; D017364:Collagenases