New experimental models for the in vitro reconstitution of human bladder mucosa.
This study describes two experimental models for the in vitro reconstitution of the human bladder mucosa (neo-bladder): human urothelial stabilized cell lines were cultured on three-dimensional matrices, collagen or platelet-fibrin gels, containing murine fibroblast 3T3-J2 cells. Low-density seeding (2x10(4) cells/ml) of both normal (TCA-48) and neoplastic cell lines (TCA-47) on collagen matrix gave rise to isolated papillar colonies, while high-density seeding (3.75x10(6) cells/ml) led to the formation of wide pluristratified epithelial sheets, resembling the normal transitional epithelium. In contrast, high-density seeding (5x10(5) cells/ml) on platelet-fibrin matrix did not allow the formation of epithelial sheets: only isolated voluminous colonies of normal TCA-48 cells, and sparse and small colonies of neoplastic TCA-47 could be observed. Growth assays and cytotoxicity reduction tests showed that the growth inhibitory effect of platelet-fibrin gel on urothelial cells was probably due to the aspecific activation of the complement contained in the plasmatic fraction, whose precipitation forms fibrin-glue. Collectively, these findings allow us to draw the following conclusions: i) neobladders obtained by culturing urothelial cells on collagen matrix reproduce normal bladder mucosa and could be utilized in pharmacological studies; and ii) platelet-fibrin gels, that specifically inhibit neoplastic urothelial cell growth, could be used as scaffolds in surgical bladder reconstitution.
De Angeli
Sergio
S
Immunotransfusional Center, Cell Culture Laboratory, Treviso Regional Hospital, I-31100 Treviso, Italy.
Del Pup
Laura
L
Fandella
Andrea
A
Di Liddo
Rosa
R
Anselmo
Giuseppe
G
Macchi
Carlo
C
Conconi
Maria Teresa
MT
Parnigotto
Pier Paolo
PP
Nussdorfer
Gastone G
GG
D016475:3T3 Cells; D000818:Animals; D001792:Blood Platelets / Q000378:metabolism; D002452:Cell Count; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D003094:Collagen / Q000378:metabolism; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods*; D005109:Extracellular Matrix / Q000378:metabolism; D005337:Fibrin / Q000378:metabolism / Q000633:toxicity; D005347:Fibroblasts / Q000166:cytology; D005782:Gels / Q000378:metabolism / Q000633:toxicity; D006801:Humans; D051379:Mice; D009092:Mucous Membrane / Q000502:physiology*; D013997:Time Factors; D018675:Toxicity Tests; D001743:Urinary Bladder / Q000166:cytology*; D001749:Urinary Bladder Neoplasms / Q000473:pathology*; D019459:Urothelium / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure
D003470:Culture Media; D005782:Gels; D005337:Fibrin; D003094:Collagen