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KMID : 0371319940460010001
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Journal of the Korean Surgical Society
1994 Volume.46 No. 1 p.1 ~ p.9
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Adult Liver Cell Culture in Hormone-Supplemented Media
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Abstract
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Liver cell culture is very attractive material for many purposes: physiologic and biochemical studies for liver, carcinogenesis of hepatoma, and research for bioartificial liver. It is more difficult to culture human adult liver cell than rat
liver
cell
because tissue of human adult liver is more stronger and activities of cell division is lower. The authors used hormone-supplemented media which contained epidermal growth factor and insulin for effective human adult liver cell culture. EGF and
insulin
have been known for effective culture of animal liver cell and increased regeneration of animal liver in vivo.
Small liver biopsies(1-3g) had been obtained from the patients with stomach cancer without liver metastasis and previous liver diseases. They were all transported to laboratory in ice-cold PBS(phosphate buffered saline) and hepatocyte were
isolated
in
HBSS(Hank's balanced salt solution) which contained 0.05% collagenase. Isolated liver cell suspension were inoculated to hormone-supplemented media and fetal bovine serum-added media respectively. And morphological aspects were observed daily
through
phase-contrast microscope.
1) The cell attachment index for hormone-supplemented media was 77% while that for fetal bovine serum-added media was 64%.
2) Monolayer of hepatocytes could be obtained 4-5 days after seeding in hormone-supplemented media, but it was not successful in fetal bovine serum-added media.
3) We could see polyhedral hepatocyte which contained multiple granules in cytoplasm and round central-located nucleolus in early phase of culture. And this typical morphology of hepatocytes were maintained for 14 days in hormone-supplemented
media,
but only 7 days in fetal bovine serum-added media. So, it was manifest that cell attachment index and cell division activity were increased and morphologic regression was slow in hormone-supplemented media.
In conclusion hormone-supplemented media which contained EGF and insulin was more effective culture media for liver cells than fetal bovine serum-added media.
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KEYWORD
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