Abstract
Epithelial cells of the gallbladder have potential to represent an important model for studies of ductal epithelia in normal and pathological states. We therefore initiated studies to establish human gallbladder epithelial cells (GBEC) in culture. GBEC were isolated by trypsinization of small tissue fragments from human gallbladders obtained at cholecystectomy; cells were plated on tissue culture dishes and grown in defined MCDB 153 medium containing added growth factors. In this medium, GBEC showed a plating efficiency of ~1%; those GBEC that attached formed colonies and proliferated, as demonstrated by autoradiographic analysis of [3H]thymidine incorporation into DNA. Cultured GBEC expressed two markers found on GBEC in situ, i.e., γ-glutamyl transpeptidase and cytokeratin 19. By using various attachment substrates, with and without added serum, increased plating efficiency and better growth were achieved. When type IV collagen was used as substrate and 10% fetal bovine serum was added to MCDB 153, passage of GBEC was possible, and cells proliferated through five to six population doublings. GBEC in culture under all conditions eventually enlarged, showed vacuolization, and demonstrated irreversible growth arrest. Nonetheless, the culture conditions described here allow for preparation of large quantities of highly enriched human GBEC.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 243-250 |
| Number of pages | 8 |
| Journal | Laboratory Investigation |
| Volume | 66 |
| Issue number | 2 |
| State | Published - 1992 |
| Externally published | Yes |
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Keywords
- Cell culture
ASJC Scopus subject areas
- Pathology and Forensic Medicine
Cite this
Biological characteristics of primary cultures of human gallbladder epithelial cells. / Hoerl, B. J.; Vroman, B. T.; Kasperbauer, Jan; La Russo, Nicholas F; Scott, R. E.
In: Laboratory Investigation, Vol. 66, No. 2, 1992, p. 243-250.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Biological characteristics of primary cultures of human gallbladder epithelial cells
AU - Hoerl, B. J.
AU - Vroman, B. T.
AU - Kasperbauer, Jan
AU - La Russo, Nicholas F
AU - Scott, R. E.
PY - 1992
Y1 - 1992
N2 - Epithelial cells of the gallbladder have potential to represent an important model for studies of ductal epithelia in normal and pathological states. We therefore initiated studies to establish human gallbladder epithelial cells (GBEC) in culture. GBEC were isolated by trypsinization of small tissue fragments from human gallbladders obtained at cholecystectomy; cells were plated on tissue culture dishes and grown in defined MCDB 153 medium containing added growth factors. In this medium, GBEC showed a plating efficiency of ~1%; those GBEC that attached formed colonies and proliferated, as demonstrated by autoradiographic analysis of [3H]thymidine incorporation into DNA. Cultured GBEC expressed two markers found on GBEC in situ, i.e., γ-glutamyl transpeptidase and cytokeratin 19. By using various attachment substrates, with and without added serum, increased plating efficiency and better growth were achieved. When type IV collagen was used as substrate and 10% fetal bovine serum was added to MCDB 153, passage of GBEC was possible, and cells proliferated through five to six population doublings. GBEC in culture under all conditions eventually enlarged, showed vacuolization, and demonstrated irreversible growth arrest. Nonetheless, the culture conditions described here allow for preparation of large quantities of highly enriched human GBEC.
AB - Epithelial cells of the gallbladder have potential to represent an important model for studies of ductal epithelia in normal and pathological states. We therefore initiated studies to establish human gallbladder epithelial cells (GBEC) in culture. GBEC were isolated by trypsinization of small tissue fragments from human gallbladders obtained at cholecystectomy; cells were plated on tissue culture dishes and grown in defined MCDB 153 medium containing added growth factors. In this medium, GBEC showed a plating efficiency of ~1%; those GBEC that attached formed colonies and proliferated, as demonstrated by autoradiographic analysis of [3H]thymidine incorporation into DNA. Cultured GBEC expressed two markers found on GBEC in situ, i.e., γ-glutamyl transpeptidase and cytokeratin 19. By using various attachment substrates, with and without added serum, increased plating efficiency and better growth were achieved. When type IV collagen was used as substrate and 10% fetal bovine serum was added to MCDB 153, passage of GBEC was possible, and cells proliferated through five to six population doublings. GBEC in culture under all conditions eventually enlarged, showed vacuolization, and demonstrated irreversible growth arrest. Nonetheless, the culture conditions described here allow for preparation of large quantities of highly enriched human GBEC.
KW - Cell culture
UR - http://www.scopus.com/inward/record.url?scp=0026583401&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026583401&partnerID=8YFLogxK
M3 - Article
C2 - 1735957
AN - SCOPUS:0026583401
VL - 66
SP - 243
EP - 250
JO - Laboratory Investigation
JF - Laboratory Investigation
SN - 0023-6837
IS - 2
ER -