Progenitor cells are responsible for formation of human prostate epithelium primary cultures.
OBJECTIVE
To analyze cell viability and morphology of primary cell cultures from CD133 immunolabeled and sorted cells from epithelium of patients suffering from benign prostate hyperplasia (BPH).


METHODS
Cells obtained from 5 patients were divided in two fractions. First fraction (CD133+/CD133-) was cultivated in DMEM with 10% FBS. Second fraction was mixed with CD133 microbeads and immunomagnetically divided into CD133+ and CD133- fractions. These cells were cultivated and followed-up for 2 weeks. Cells were stained for Annexin V FITC/propidium iodide.


RESULTS
Seventy CD133+/CD133- cultures, thirty-one of CD133+ and thirty-one of CD133- cells were established. There were 5-fold and 3-fold increase of CD133+/CD133- and CD133+ cell number after 2 weeks, respectively. CD133+/CD133- and CD133+ monolayers displayed epithelial-like morphology and cytokeratine expression. CD133- cultures collapsed. Cell viability within CD133+ and CD133- populations was 90.1-/+6.3% and 24.3-/+6.2%, respectively. Apoptotic index was 9.0-/+6.1% and 28.5-/+23.8% within CD133+ and CD133- cultures, respectively.


CONCLUSIONS
CD133 separated human primary epithelial cell cultures displayed differences in morphology, viability and apoptosis occurrence. Immunomagnetic sorting can be recommended in each in vitro experiments with primary cell cultures in order to provide more objective results.
Department of Urology, Nicolaus Copernicus University (NCU), Bydgoszcz, Poland. tomaszdrewa@wp.pl
T
T
Drewa
Z
Z
Wolski
L
L
Pokrywka
R
R
Debski
J
J
Styczynski
D017209:Apoptosis; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002470:Cell Survival; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology*; D004848:Epithelium / Q000473:pathology*; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D008297:Male; D009336:Necrosis; D011467:Prostate / Q000166:cytology* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology*