TY - JOUR
T1 - Long-term culture of porcine bladder epithelial cells
AU - Ehmann, Ursula K.
AU - Terris, Martha K.
N1 - Funding Information:
We thank Dr, Luis Fajardo in whose laboratory most of the experiments were performed. We appreciate the help of Ms. Bonnie Bell in obtaining and processing the primary bladder tissues and of Ms. Lusijah Rott for operation of the FACS. Dr. Raymond Sobel graciously contributed some of the supplies tor this project. This work was supported by the Department of Veterans Affairs and by the Stanford Office of the Dean of Research, OTL Research Incentive Award.
PY - 2002
Y1 - 2002
N2 - Epithelial cells from normal pig bladders proliferated when cocultured with lethally irradiated feeder cells of the LA7 rat mammary tumor line. When the bladder cells and feeders were plated together r at a confluent density, the bladder cells proliferated as the feeder cells died, resulting in a confluent culture of bladder cells. The bladder cells were successfully subcultured by plating with freshly irradiated LA7 feeder cells. In this way, bladder cells from five pigs were carried to confluency in passages 1, 4, 7, 7, and 13, amounting to at least 6, 18, 24, 26, and 45 doublings in culture, respectively, and none showed signs of slowed proliferation at the time of culture termination. Fibroblasts never became a prominent feature of these cultures, and their frequency was determined to be about 26 fibroblasts per 105 cells in passage 9. Pig bladder cells in 0,5% serum doubled in number in slightly over 3 d, whereas cells in 5,0% serum doubled in about 6 d. In fresh medium without feeder cells only minimal proliferation of bladder cells occurred. In LA7-conditioned medium the bladder cell numbers decreased, leading to the conclusion that the stimulus from LA7 cells is mechanically or physically transmitted. The bladder cells reacted with antibodies to keratins 7 and 18 but not to keratin 14 or vimentin. Tight junctions, visualized with an antibody to the ZO1 protein, connected all the cells to their neighbors. Most cells in passage 9 carried the diploid chromosome number of 38.
AB - Epithelial cells from normal pig bladders proliferated when cocultured with lethally irradiated feeder cells of the LA7 rat mammary tumor line. When the bladder cells and feeders were plated together r at a confluent density, the bladder cells proliferated as the feeder cells died, resulting in a confluent culture of bladder cells. The bladder cells were successfully subcultured by plating with freshly irradiated LA7 feeder cells. In this way, bladder cells from five pigs were carried to confluency in passages 1, 4, 7, 7, and 13, amounting to at least 6, 18, 24, 26, and 45 doublings in culture, respectively, and none showed signs of slowed proliferation at the time of culture termination. Fibroblasts never became a prominent feature of these cultures, and their frequency was determined to be about 26 fibroblasts per 105 cells in passage 9. Pig bladder cells in 0,5% serum doubled in number in slightly over 3 d, whereas cells in 5,0% serum doubled in about 6 d. In fresh medium without feeder cells only minimal proliferation of bladder cells occurred. In LA7-conditioned medium the bladder cell numbers decreased, leading to the conclusion that the stimulus from LA7 cells is mechanically or physically transmitted. The bladder cells reacted with antibodies to keratins 7 and 18 but not to keratin 14 or vimentin. Tight junctions, visualized with an antibody to the ZO1 protein, connected all the cells to their neighbors. Most cells in passage 9 carried the diploid chromosome number of 38.
KW - Bladder
KW - Cell proliferation
KW - Culture method
KW - Feeder cells
KW - Juxtacrine
KW - Transitional epithelial cells
UR - http://www.scopus.com/inward/record.url?scp=0036253453&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036253453&partnerID=8YFLogxK
U2 - 10.1290/1071-2690(2002)038<0137:LTCOPB>2.0.CO;2
DO - 10.1290/1071-2690(2002)038<0137:LTCOPB>2.0.CO;2
M3 - Article
C2 - 12026161
AN - SCOPUS:0036253453
SN - 1071-2690
VL - 38
SP - 137
EP - 141
JO - In Vitro Cellular and Developmental Biology - Animal
JF - In Vitro Cellular and Developmental Biology - Animal
IS - 3
M1 - 3803137
ER -