Introduction and Objectives
The CD133 surface antigen is considered to be a marker stem cells and is used to isolate prostatic epithelial stem cells. This subpopulation is probably playing an important role in prostate cancer and BPH, as well. The prostate epithelial CD133 positive cells isolation method and our results are presented.
Material and Methods
Prostate epithelial cells were isolated from the material from 19 adenomectomies performed in two centers. Time of transportation was 1 h in 13 cases and 24 h in 6 cases. Tissue was transported in cold medium with antibiotics. Patients operated near lab aged 56 to 80 (68.1±7.9) while patients operated far away from lab aged 71 to 83 (76.5±4.9). Tissue was cut into 1mm pieces and incubated for 6 h in collagenase type I (1 mg/ml, Sigma). Cells were washed, resuspended and counted. CD133+ cells were isolated using magnetically labelled anti-CD133 antibodies (Miltenyi Biotec). Cells were cultivated in 25 cm2 T-flasks in medium designed for prostate epithelial cells.
Results
Total numbers of living cells obtained after specimens enzymatic digestion from both centers were comparable (0.6–6.0x106). The percentage of CD133 positive cells was 4.9±1.6 of total living cells obtained from center near lab (1 h of transportation time). The percentage of CD133 positive cells was 3.6±2.8 of total living cells obtained from center far away form lab (24 h of transportation time).
Conclusions
Tissue transportation time even up to 24 hours did not influence on epithelial cell isolation procedure and total epithelial cell viability. It seems that patient age can influence on stem cell number within prostate epithelium.
Article info
Identification
Copyright
© 2009 European Association of Urology. Published by Elsevier Inc. All rights reserved.
