Differentiation of hepatic oval cell into mature hepatocyte induced by hepatic stellate cells / 中华肝脏病杂志
Chinese Journal of Hepatology
;
(12): 765-770, 2009.
Article
Dans Chinois
| WPRIM
| ID: wpr-306677
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of hepatic stellate cells in the differentiation of hepatic oval cells into adult hepatocyte.</p><p><b>METHODS</b>The oval cell were cocultured with primary hepatic stellate cells (HSC) in the same well (M-coculture) or separately cultured with HSC by millIcell (S-coculture). Oval cells were cultured alone as control; the expression of adult hepatocyte marker HNF-4alpha, albumin, and oval cell marker AFP, CK-19 in each group were detected by real-time PCR and western-blot. Phenotype changes were observed by transmission electron microscope (TEM); PAS staining was used to detect the quantity of glycogen granule in oval cell. Albumin level in supernatant was detected using ELISA kit.</p><p><b>RESULT</b>(1) The relative level of HNF-4alpha and albumin mRNA expression compared with pre-coculture M-coculture HNF-4a 1.9+/-0.2, 10.7+/-1.2, 12.0+/-1.3; albumin 5.7+/-1.6, 110.7+/-13.7, 173.6+/-22.3. S-coculture 1.4+/-0.1, 3.2+/-0.6, 8.9+/-1.4 times; albumin 2.9+/-1.4, 22.3+/-8.5, 96.3+/-16.3. The relative level of HNF-4a and albumin mRNA expression in coculture group (M- and S-coculture) were higher than control group (LSD-t 32.98, 10.08, 13.38, 7.96; P less than 0.01); and a higher level of HNF-4a and albumin was found in M-coculture group compared to S-coculture group (LSD-t 32.98, 25.65; P less than 0.01). The relative level of AFP and CK-19 mRNA expression compared with pre-coculture M-coculture 1.1+/-0.2, 0.2+/-0.0, 0.0+/-0.0; S-coculture group AFP 1.0+/-0.2, 0.2+/-0.1, 0.1+/-0.0; CK-19 0.6+/-0.1, 0.1+/-0.0, 0.0+/-0.0; control group AFP 1.0+/-0.1, 1.0+/-0.1, 1.1+/-0.1, CK-19 1.0+/-0.1, 1.1+/-0.1, 1.0+/-0.1. The relative level of AFP and CK-19 mRNA expression in coculture group (M- and S-coculture) were lower than that in control group (LSD-t 37.99, 34.50, 13.59, 22.46; P less than 0.01). (2) The albumin secretion was detected in M-coculture 14 day (15.30+/-0.09) ng/ml, 21 (20.98+/-0.12) ng/ml; S-coculture 14 day (11.41+/-0.13) ng/ml, 21 day(15.12+/-0.17) ng/ml. (3) It showed more organelles such as endoplasmic reticulum, mitochondrion and Golgi apparatus in oval cells cocultured with HSC. And cholangiole-like structure appeared between oval cells cocultured with HSC. (4) PAS staining showed glycogen granules could be observed in coculture groups.</p><p><b>CONCLUSION</b>HSC can induce differentiation of oval cell into mature hepatocyte.</p>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Cellules souches
/
ARN messager
/
Alphafoetoprotéines
/
Différenciation cellulaire
/
Cellules cultivées
/
Réaction de polymérisation en chaîne
/
Rat Sprague-Dawley
/
Microscopie confocale
/
Techniques de coculture
/
Hépatocytes
Limites du sujet:
Animaux
langue:
Chinois
Texte intégral:
Chinese Journal of Hepatology
Année:
2009
Type:
Article
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