Successful mouse hepatocyte culture with sandwich collagen gel formation / 대한외과학회지
Journal of the Korean Surgical Society
; : 202-208, 2013.
Article
en En
| WPRIM
| ID: wpr-160123
Biblioteca responsable:
WPRO
ABSTRACT
PURPOSE: Primary mammalian hepatocytes largely retain their liver-specific functions when they are freshly derived from donors. However, long-term cultures of functional hepatocytes are difficult to establish. To increase the longevity and maintain the differentiated functions of hepatocytes in primary culture, cells can be cultured in a sandwich configuration of collagen. In such a configuration, hepatocytes can be cultured for longer periods compared with cultures on single layers of collagen. However, research regarding mouse hepatocytes in sandwich culture is lacking. METHODS: Primary mouse hepatocytes were sandwiched between two layers of collagen to maintain the stability of their liver-specific functions. After gelation, 2 mL of hepatocyte culture medium was applied. RESULTS: After 24 hours, 5, 10 days of culture, the collagen gel sandwich maintained the cellular border and numbers of bile canaliculi more efficiently than a single collagen coating in both high and low density culture dishes. Reverse transcription-polymerase chain reaction analysis of alpha-1-antitrypsin (AAT), hepatocyte nuclear factor 4 alpha (HNF4A), alphafetoprotein, albumin, tryptophan oxygenase (TO), the tyrosine aminotransferase gene, glucose-6-phosphatase, glyceraldehyde-3-phosphate dehydrogenase for mouse primary hepatocytes cultured on collagen coated dishes and collagen gels showed superior hepatocyte-related gene expression in cells grown using the collagen gel sandwich culture system. AAT, HNF4A, albumin, TO were found to be expressed in mouse hepatocytes cultured on collagen gels for 5 and 10 days. In contrast, mouse hepatocytes grown on collagen-coated dishes did not express these genes after 5 and 10 days of culture. CONCLUSION: The collagen gel sandwich method is suitable for primary culture system of adult mouse hepatocytes.
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Índice:
WPRIM
Asunto principal:
Oxidorreductasas
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Donantes de Tejidos
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Triptófano Oxigenasa
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Tirosina Transaminasa
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Canalículos Biliares
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Expresión Génica
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Colágeno
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Glucosa-6-Fosfatasa
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Hepatocitos
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Factor Nuclear 4 del Hepatocito
Límite:
Animals
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Humans
Idioma:
En
Revista:
Journal of the Korean Surgical Society
Año:
2013
Tipo del documento:
Article