Culture of pancreatic duct-derived stem cells of rats in vitro in the dynamic three-dimension cell culture system
10.3760/cma.j.issn.0254-1785.2012.06.014
- VernacularTitle:动力性三维细胞培养系统建立大鼠胰腺导管来源干细胞系的研究
- Author:
Xuchun CHEN
;
Ying CHENG
;
Guichen LI
;
Rui SHI
;
Xiaohang LI
;
Da LI
;
Yongfeng LIU
- Publication Type:Journal Article
- Keywords:
Rats;
Pancreatic ducts;
Stem cells;
Cell culture techniques
- From:
Chinese Journal of Organ Transplantation
2012;33(6):371-375
- CountryChina
- Language:Chinese
-
Abstract:
Objective To isolate,purify and identify pancreatic duct derived stem cells (PDSCs) from the pancreatic duct of rats,and culture in the three-dimension cell culture system.Methods Adult male Wistar rats underwent perfusion with collagenase V via the pancreatic duct,then the pancreas was surgically procured,digested,followed by discontinued density gradient centrifuge to isolate ductal tissue from islets.The acinar and ductal tissue was cultivated in serumcontaining medium in the three-dimension cell culture to obtain adherent cells,as PDSCs,which were expanded by consecutive passages.The morphology and characterization of PDSCs on phenotype were examined.Results PDSCs could be obtained through in situ collagenase V digestion,discontinued density gradient centrifuge,and culture in the three dimension cell culture system.Morphologically,PDSCs had remarkable size,most with one nucleus.PDSCs grew in many layers in three-dimension cell culture system.PDSCs was revealed to express CD29,CD73,CD90,CD105,but not CD14,CD19,CD34,CD45 by FACS,in agreement with MSCs.Conclusion PDSCs of rats could be obtained through in situ collagenase V digestion,discontinued density gradient centrifuge,and culture in the three-dimension cell culture system.PDSCs lines were successfully established.
