Expression of stem cell markers CK-19 and PDX-1 mRNA in pancreatic islet samples of different purity from rats.
BACKGROUND
Islet stem cells are more or less retained in the procedure of islet isolation and purification, and are transplanted together with islet grafts. Keratoprotein (CK-19) and pancreatic duodenal hox gene 1 (PDX-1) are markers of stem cells. This study was undertaken to examine the expression of these markers in pancreatic islet samples of different purity from rats.


METHODS
A total of 30 male Sprague-Dawley rats were randomly assigned to 3 groups to undergo perfusion with V-type collagenase via the pancreatic duct, then the pancreas was excised, diced, shaken, digested and centrifuged to obtain islet sediments. The sediment from group A was not purified, while that from group B was purified with 25% Ficoll-400 and that from group C with 25% and 11% Ficoll-400. RNA was extracted from the different islet samples for reverse transcriptase-polymerase chain reaction (RT-PCR). The expression of the pancreatic stem cell markers CK-19 and PDX-1 was assessed.


RESULTS
The purity of islets in samples was (43.6+/-6.29)% in group A; (65.3+/-4.40)% in group B; and (77.6+/-6.36)% in group C (P<0.05). The expression of CK-19 and PDX-1 mRNA was significantly higher in group A than in groups B and C, but group C showed the lowest level of expression.


CONCLUSIONS
The expression of CK-19 and PDX-1 mRNA in islet samples of different purity suggests the presence of stem cells in all islet samples.
Yang
Chuang
C
Department of Hepatobiliary Surgery, Mianyang Third People's Hospital, Mianyang 621000, China.
Wang
Ji-Ming
JM
Du
Cheng-You
CY
Xue
Dong
D
D000818:Animals; D004195:Disease Models, Animal; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000096:biosynthesis / Q000235:genetics*; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D053539:Keratin-19 / Q000096:biosynthesis / Q000235:genetics*; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics*
D018398:Homeodomain Proteins; D053539:Keratin-19; D012333:RNA, Messenger; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein