Isolation, cultivation, and identification of human fibroblasts from cirrhotic liver
10.3760/cma.j.issn.1673-4203.2017.04.008
- VernacularTitle:肝硬化组织来源纤维母细胞的分离、培养及鉴定
- Author:
Yalei ZHAO
;
Kaiming ZHANG
;
Yuling SUN
;
Wenqi LI
;
Yanpeng ZHANG
;
Yipu ZHAO
- Keywords:
Liver cirrhosis;
Myofibroblasts;
Liver;
Tissue block method
- From:
International Journal of Surgery
2017;44(4):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a method for isolating and culturing fibroblasts from cirrhotic liver in vitro,observe the biological characteristics of fibroblasts.Methods Human fibroblasts from cirrhotic liver were isolated and cultured by adhering to the culture plastic.The morphologic and growth characteristics of the acquired cells were observed by microscopy imaging and cell counting.The expression of FSP-1 and Vimentin of fibroblasts was detected through immunofluorescence and western blotting.Results The tissue blocks were well-adherent to the culture plastic within 2 hours.Hepatocyte-like cells were observed surrounding the blocks within about 1 week,and spindle-shaped fibroblasts were observed within about 2 weeks.After a series of passage,the attached cells became proliferated quickly.The growth curves of the passage 3,4 and 5 were quite similar.The result of immunofluorescence showed the expression of FSP1 and Vimentin was positive.The positive rate was respectively (90.6 ± 1.0) ％ and (91.2-± 4.1) ％.Conclusions Human fibroblasts from cirrhotic liver can be cultured successfully through adherent property.The method for isolation and culture of fibroblasts from cirrhotic liver is convenient,efficient,stable and cultured cells remain naive biological characteristics.
