In vitro isolation, culture and differentiation of nestin positive cells in the pancreas of newborn rats
- VernacularTitle:新生大鼠胰腺nestin阳性细胞的体外分离、培养和分化的研究
- Author:
Jingqi TIAN
;
Rixing BAI
;
Haiyan ZHANG
;
Weihong LI
;
Libo SUN
;
Jianzhen JIE
;
Deheng ZHANG
- Publication Type:Journal Article
- Keywords:
Pancreas;
Stem cells;
Islet of Langerhans;
Cell culture
- From:
Chinese Journal of General Surgery
2000;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the isolation, culture of nestin positive cells of the pancreas in newborn rats. Methods The whole pancreas of neonatal rats were digested with collagenase, followed by incubation under pH 7. 6 serum RPMI 1640 for 24 - 36 h and then under pH 7. 4 serum free RPMI 1640 (bFGF.EGF 20 ng/ml.1% N2) for 18-24 d. The expression of insulin.glucagon and nestin were detected by immunofluorescence technique. Nestin.CK19 were detected by RT-PCR. Resides Cells attached to the surface of plates after 24 h incubation under pH 7. 6 condition, and islet-like masses were obtained after 18 -24 d incubation. A monolayer of cells grew out after 24 h of passage of islet-like masses. Nestin positive cells was detected after 24 - 36 h incubation, with no expression of insulin and glucagon. Positive cells of insulin and glucagon were detected in islet-like masses after 24 h passage. Nestin positive cells were detected by RT-PCR in islet-like masses after 24 h passage, but no CK19. Conclusion Insulin and glucagon were expressed in islet-like masses after passage. Nestin positive cells in the pancreas of neonatal rats possessed character of islet stem cells.
