ISOLATION AND CHARACTERIZATION OF BILIARY EPITHELIAL-CELLS FROM RAINBOW-TROUT LIVER

被引:2
作者
BLAIR, JB
OSTRANDER, GK
MILLER, MR
HINTON, DE
机构
[1] OKLAHOMA STATE UNIV, DEPT BIOCHEM & MOLEC BIOL, DIV AGR SCI & NAT RESOURCES, STILLWATER, OK 74078 USA
[2] OKLAHOMA STATE UNIV, DEPT ZOOL, ENVIRONM TOXICOL PROGRAM, STILLWATER, OK 74078 USA
[3] W VIRGINIA UNIV, HLTH SCI CTR, DEPT BIOCHEM, MORGANTOWN, WV 26505 USA
[4] UNIV CALIF DAVIS, SCH VET MED, DEPT MED, DAVIS, CA 95616 USA
关键词
TROUT; LIVER; HEPATOCYTES; BILIARY CELLS; FISH; CELL SEPARATIONS; LECTIN BINDING; GLYCOLIPIDS;
D O I
暂无
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Lectin binding and density gradient centrifugation were explored for isolating epithelial cells from trout liver. Hepatocytes exhibited preferential attachment to coverslips coated with Phaseolus vulgaris erythroagglutinin. Biliary epithelial cells attached with glycine max agglutinin; however, significant attachment of cellular debris limited the use of glycine max agglutinin. Percoll-density gradient centrifugation separated liver cells into two distinct populations with biliary cells and hepatocytes banding at densities of 1.04 and 1.09, respectively. A discontinuous gradient composed of 13% Ficoll (wt/wt) separated biliary cells from hepatocytes. The recovery of highly enriched biliary epithelial cells from trout liver using Ficoll gradients yielded approximately 8 million cells (0.1 ml packed cells) from 10 g liver. Western blot analysis demonstrated that the cytokeratin profile for extracts from biliary epithelial cell-enriched populations differ significantly from those seen with whole liver extracts or with extracts from hepatocyte-enriched populations. Ficoll-gradient purified biliary cells and hepatocytes attached to culture plates coated with trout skin extract and carried out linear incorporation of leucine into protein and thymidine into DNA for 24 h. A mixture of growth hormones (insulin, epidermal growth factor, and dexamethasone) stimulated thymidine incorporation into DNA; however, long-term culture of dividing biliary epithelial cells was not achieved. Chemical analysis of neutral and acidic glycolipids indicated that hepatocytes and biliary cells have similar glycolipid profiles with an exception in the region of GM3 mobility, which is attributable to differences in the ceramide moiety. These studies provide a starting point for further characterization of unique cell types of the trout liver that may be important in their response to toxic and carcinogenic agents.
引用
收藏
页码:780 / 789
页数:10
相关论文
共 50 条
[1]   THE STEM-CELLS OF THE LIVER - A SELECTIVE REVIEW [J].
ATERMAN, K .
JOURNAL OF CANCER RESEARCH AND CLINICAL ONCOLOGY, 1992, 118 (02) :87-115
[2]   PATHOBIOLOGY OF CHEMICAL HEPATOCARCINOGENESIS - RECENT PROGRESS AND PERSPECTIVES .1. CYTOMORPHOLOGICAL CHANGES AND CELL-PROLIFERATION [J].
BANNASCH, P .
JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, 1990, 5 (02) :149-159
[3]   ISOLATED TROUT LIVER-CELLS - ESTABLISHING SHORT-TERM PRIMARY CULTURES EXHIBITING CELL-TO-CELL INTERACTIONS [J].
BLAIR, JB ;
MILLER, MR ;
PACK, D ;
BARNES, R ;
TEH, SJ ;
HINTON, DE .
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY, 1990, 26 (03) :237-249
[4]   ULTRASTRUCTURAL ALTERATIONS IN LIVER OF MEDAKA (ORYZIAS-LATIPES) EXPOSED TO DIETHYLNITROSAMINE [J].
BRAUNBECK, TA ;
TEH, SJ ;
LESTER, SM ;
HINTON, DE .
TOXICOLOGIC PATHOLOGY, 1992, 20 (02) :179-196
[5]   THE IMMUNOCYTOCHEMISTRY OF CYTOKERATIN IN FISH-TISSUES [J].
BUNTON, TE .
VETERINARY PATHOLOGY, 1993, 30 (05) :418-425
[6]   MITOGENIC ACTIVITY FROM TROUT EMBRYOS [J].
COLLODI, P ;
BARNES, DW .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (09) :3498-3502
[7]  
COUCH JA, 1985, ENVIRON CARCINOG REV, V3, P63
[8]  
COUCH JA, 1987, JNCI-J NATL CANCER I, V79, P297
[9]  
Farber E., 1992, ROLE CELL TYPES HEPA, P1
[10]  
FAUSTO N, 1992, ROLE CELL TYPES HEPA, P89