Isolation And Biological Behavior Study Of CD133Positive Cells From Bladder Cancer Cell Line5637

Object：To isolate and culture the CD133Positive cells from bladdercancer cell line5637, to assess their invasiveness, tumorigenic ability, and toexplore whether the CD133-positive human bladder cancer cell have the tumorstem cells properties.Methods：Study of the expressions of CD133in bladder cancer cell line5637by flow cytometry，Applied with the magnetic cell sorting(MACS)technology, we purified CD133positive cells from bladder cancer cell line5637and identified by flow cytometry. Identification of proliferation,Clonogenic capacity and migration capacity of CD133~+and and CD133~-cellsby MTT assay, Flat colony formation，Wound healing assay；Identification ofdifferentiation by flow cytometer.Results：1CD133~+was expressed on a small fraction of cells in bladder cancercell line5637, the percentage of CD133~+cells was1.45%Detection ofexpression of CD133in bladder cancer cells line5637by flow cytometer.2CD133~+cells purified by MACS were in a considerable purity of93.42%. Purity examination of CD133~+cells by flow cytometer.3The results of MTT assay, Flat colony formation，Wound healing assayshow that CD133~+cells display higher proliferative potential,clonogeniccapacity and migratory capacity than CD133_cells.4After cultured in serum-containing medium,CD133~+cells coulddifferentiate into a different phenotype of CD133~-tumor cells, the percentageof CD133~+cell population decreased rapidly as detecded by flow cytometer. Conclusion：1Detection of expression of CD133in bladder cancer cell line5637byflow cytometry.2CD133~+cells could be effectively obtained high-speed by MACS；CD133~+cells sorted by flow cytometer were in a considerable purity of93.45%and can be used for the follow-up experiment3CD133~+cells displayed stem cell-like properties of self-renewal anddifferentiation compared with the CD133~-cells in vitro. CD133~+cells posessedstronger motility than CD133~-cells in vitro, CD133~+cells may contribute tothe recurrence and metastasis of bladder carcinoma cell.