Long term organ culture of human prostate tissue in a NASA-designed rotating wall bioreactor.
OBJECTIVE
To maintain ex vivo integral prostatic tissue including intact stromal and ductal elements using the NASA-designed Rotating Wall Vessel (RWV) which maintains colocalized cells in an environment that promotes both three-dimensional cellular interactions together with the uniform mass transfer of nutrients and metabolic wastes.


METHODS
Samples of normal prostate were obtained as a byproduct of transurethral prostatectomy or needle biopsy. Prostatic tissue dissected into small 1 x 1 mm. blocks was cultured in the Rotating Wall Vessel (RWV) Bioreactor for various time periods and analyzed using histological, immunochemical, and total cell RNA assays.


RESULTS
We report the long term maintenance of benign explanted human prostate tissue grown in simple culture medium, under the simulated microgravity conditions afforded by the RWV bioreactor. Mesenchymal stromal elements including blood vessels and architecturally preserved tubuloglandular acini were maintained for a minimum of 28 days. Cytokeratins, vimentin and TGF-beta2 receptor and ligand were preserved through the entire culture period as revealed by immunocytochemistry. Prostatic acid phosphatase (PAP) was continuously expressed during the culture period, although somewhat decreased. Prostatic specific antigen (PSA) and its transcript were down regulated over time of culture. Prostatic carcinoma cells from the TSU cell line were able to invade RWV-cultured benign prostate tissue explants.


CONCLUSIONS
The RWV bioreactor represents an additional new technology for culturing prostate tissue for further investigations concerning the basic physiology and pathobiology of this clinically important tissue.
NIH/NASA Center for Three Dimensional Tissue Culture, Laboratory of Cellular and Molecular Biophysics, National Institute for Child Health and Human Development, Bethesda, Maryland, USA.
L
L
Margolis
S
S
Hatfill
R
R
Chuaqui
C
C
Vocke
M
M
Emmert-Buck
W M
WM
Linehan
P H
PH
Duray
D019149:Bioreactors; D046508:Culture Techniques / Q000295:instrumentation*; D004867:Equipment Design; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D009361:Neoplasm Invasiveness; D011467:Prostate / Q000737:chemistry / Q000166:cytology; D011471:Prostatic Neoplasms / Q000473:pathology; D012333:RNA, Messenger / Q000032:analysis; D013997:Time Factors; D014407:Tumor Cells, Cultured; D014481:United States; D018557:United States National Aeronautics and Space Administration
D012333:RNA, Messenger