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BSP-11
(P)
GENETIC STABILITY IN IN VITRO CULTURED HUMAN UROTHELIAL CELLS
Clara Ibel CHAMORRO JIMENEZ
1
, Said ZEIAI
2
, Johanna LUNDIN
3
, Gisela REINFELDT ENGBERG
4
, Johana FERNANDEZ
5
,
Agneta NORDENSKJÖLD
6
and Magdalena FOSSUM
7
1) Karolinska Institutet, Women and Children Health, Stochkolm, SWEDEN - 2) Karolinska University Hospital,
Department of Pediatric Surgery, Astrid Lindgren Children's Hospital, Stockholm, SWEDEN - 3) Karolinska Institutet,
Department of Women's and Children's Healt, Stochkolm, SWEDEN - 4) Karolinska University Hospital, Department of
Women's and Children's Health and Center of Molecular Medicine, Stochkolm, SWEDEN - 5) Karolinska Institutet,
Department of Women's and Children's Health, Stochkolm, SWEDEN - 6) Karolinska University Hospital, Pediatric
Surgery, Unit of Urology, Astrid Lindgren Children's Hospital, Stochkolm, SWEDEN - 7) Karolinska University Hospital,,
Pediatric Surgery, Unit of Urology, Astrid Lindgren Children's Hospital, Stochkolm, SWEDEN
PURPOSE
Cultured human urothelial cells have a potential use in regenerative medicine for treatment of different urological
conditions. We predict that high quality control standards will be imperative for all cellular therapy products. In this
study we aimed to investigate genomic stability in human urothelial cells in long-term culture to better understand the
effects of in vitro propagation
MATERIAL AND METHODS
The Affymetrix, human genome U133 2 plus set (HG-U133) was used to compare gene expression profiles at early and
late passage (p2 vs p8). The Ingenuity Pathway Knowledge database and biological function software tools were used to
interpret gene expression profiles and to identify upstream regulators.
Chromosomal structural instability was investigated with high resolution Comparative Genomic Hybridization analysis
(CGH) with special focus on oncogenic regions
RESULTS
Bioinformatics analysis of the differentially expressed genes identified several upstream transcriptional regulators and its
target molecules. These regulators play an important role in controlling cell-fate and are frequently associated with
changes in the cell cycle. Long-term cultured cells had a normal expression pattern of oncogenes/tumor suppressor
genes and also of genes associated with bladder cancer. Furthermore, CGH analysis indicated no gain or loss of
chromosomal regions of bladder cancer related genes or any other genome regions down to 20kb.
CONCLUSIONS
Urothelial cells in vitro are stable at a submicroscopic genetic level. Patterns of gene expression demonstrate a decrease
in proliferative capacity and no increase in oncogenic activity after long-term culture. We suggest array CGH and gene
expression studies on selected genes as a routine for regenerative cell therapies.