Reconstituted basement membrane promotes morphological and functional differentiation of primary human prostatic epithelial cells

Abstract

Prostatic epithelial cells undergo rapid proliferation and lose their ability to synthesize and secrete prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) under standard tissue culture conditions. Herein, we compared the morphology, growth, secretory activity, and intermediate filament expression of human prostatic epithelial cells cultured on either standard tissue culture plastic or reconstituted basement membrane. Epithelial cells grown on plastic exhibited a 10-fold increase in proliferation and a higher percentage of cells in the S-phase of the cell cycle compared to cells cultured on basement membrane. However, cells grown on basement membrane secreted markedly higher levels of PSA and PAP. The basement membrane-induced enhancement of secretory activity was potentiated by dihydrotestosterone (DHT) and prostate stromal cell conditioned medium. Morphological studies showed that cells plated on basement membrane formed organoid-like clusters and maintained several aspects of differentiated epithelium including abundant secretory vesicles, microvilli, and desmosomes with associated cytoskeletal elements. Cultivation of epithelial cells on basement membrane components also suppressed the expression of vimentin, a mesenchymal intermediate filament polypeptide. However, cytokeratin expression was abnormal in cells grown on either surface. These results indicate that the differentiated properties of prostatic epithelial cells are promoted by cultivation on reconstituted basement membrane in the presence of DHT and stromal cell conditioned medium.

Cell–cell interaction in prostate gene regulation and cytodifferentiation

“…The suspension was passed the next day through a 70-m mesh nylon filter (Falcon 2350, Becton Dickinson) and then aspirated with a 23-gauge needle. Prostatic epithelial cells were separated from the stromal cells by centrifugation in a Percoll density gradient (14). Two 10-ml iso-osmotic Percoll (Pharmacia) solutions were prepared as follows: (i) ϭ 1.07: 5 ml Percoll, 0.9 ml 1.5 M NaCl, 4.1 ml H 2 O; and (ii) ϭ 1.035: 2.3 ml Percoll, 0.9 ml 1.5 M NaCl, 6.8 ml H 2 O.…”

Epithelial-Restricted Gene Profile of Primary Cultures from Human Prostate Tumors: A Molecular Approach to Predict Clinical Behavior of Prostate Cancer

“…Each normal epithelial cell type has a specific pattern of cytokeratin expression, which can be used as a differentiation marker (19). Cytospins of cultured cells were analyzed by immunocytochemistry using antibodies against several common prostate epithelial or stromal markers.…”

Epithelial stem cells in human prostate growth and disease

“…43 Matrigel has been widely used to study epithelial differentiation in breast epithelial cell lines 44 and in prostate. 45 Since one of the definitions of a stem cell is pluripotency, the ability to produce different cell types, we used Matrigel to examine the capacity of our putative stem-cell population to fully differentiate. 41 Cells that had been grown for 14 days, as described above, were harvested and cells derived from single type II colonies were cultured on Matrigel in the presence of androgen and stromal cell-conditioned medium.…”