Isolation and culture of hepatic oval cells and in vivo transplantation in mice
Date Issued
2003-07-31
Date
2003-07-31
Author(s)
袁瑞晃
DOI
912314B002250
Abstract
Stem cells with multiple differentiation
potential have been verified in several
different tissues. The existence of a liver
stem cell was controversial for many years,
but it is now generally accepted that liver
contains kinds of cells with stem-like
properties and these cells can be activated
under certain conditions and differentiated
into hepatocytes. By reason of their oval
shaped nucleus and scanty cytoplasm, these
immature epithelial cells were called “oval
cells”. Even though primary culture of the
hepatocytes had been done for years, the
3
impacts on proliferative ability and viability
after several generations made their usage
limited. Under a variety of pathological
conditions, similar small cells can be found
in human liver too. Therefore, searching for
the hepatic stem cells that can be cultured for
a long period and application of these cells in
hepatic cell transplantation become more and
more imminent. In this study, we use
chemical agent combined with partial
hepatectomy as a model of diseased liver to
activate hepatic oval cells for isolation and
cell culture. Then these cells will be
transplanted into the diseased mice in order
to verify the ability of differentiation and
repopulation of oval cells in vivo.
Both C57BL/6 and DPPIV (Dipeptidyl
Peptidase IV) knock-outmice will be used in
this study. C57BL/6 mice will receive oral
gavage of Acetylaminofluorene(AAF,
7.5mg/kg/day, dissolved in polyethylene
glycol) for 14 days totally. Two-third partial
hepatectomy will be performed at the 24
hours after the first six daily gavages (day
7th). Hepatic oval cells will be isolated using
two-step collagenase perfusion method and
seeded in collagen-coated tissue culture
dishes. Immunohistochemical studies about
the γ-glutamyl transpeptidase、
α-fetoprotein、albumin、cytokeratin 7、
cytokeratin 18、cytokeratin 19、Thy-1 will be
used to evaluated the characters of these cell..
Then the oval cells will be transplanted into
AAF pretreated PPIV knockout mice
immediately after two-third partial
hepatectomy. The recipients will be
sacrificed two weeks, four weeks, two
months, four months and six months after
the cell transplantation. Histochemical
staining will be used to distinguish the donor
cells and albumin、cytokeratin 7、Thy-1 will
be used to identify the character of oval cell
proliferation and differentiation.
Subjects
oval cell isolation
oval cell
transplantation
transplantation
partial hepatectomy
SDGs
Publisher
臺北市：國立臺灣大學醫學院外科
Type
report
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