Canine urinary bladder epithelial cells: Preparation for cell culture by enzyme dispersion

Bonar, R. A.; Reich, Charles F.; Sharief, Yousuf

doi:10.1007/bf00256846

Cited by 16 publications

(9 citation statements)

References 18 publications

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“…These observations are quite distinct from those of others, who described the mass culture of urothelial cells as monolayers growing on plastic substrate (Stone et al, 1975; Berky & Zoloter, 1977; Bonar, Reich & Sharlef, 1977; Rozell, Douglas & Irving, 1977;Kirk et al, 1985). Most recently, Kirk et al (1985) has reported that adult human urothelial cells can be grown as monolayers in serum-free MCDB 152 medium supplemented with a variety of additions including bovine pituitary extract, low Ca++ ( M), insulin, hydrocortisone and epidermal growth factor.…”

contrasting

confidence: 59%

Long term growth and maintenance of stratified rat urothelium in vitro

1989

Cell Proliferation

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Culture conditions that allow long term growth and maintenance of rat urothelium have been determined using short (3 to 8 days) and long (14 to 60 days) term measurements of cell density and tritiated thymidine incorporation as indices. The basal nutrient medium utilized was a mixture of 199 plus Ham's F 12 (1:1) supplemented with insulin (1 μ/ml) and hydrocortisone (1 μ/ml). Long term culture of urothelium seems to require porous collagen. Porous albumin, or plastic dishes thinly coated with albumin, collagen, fibronectin or mixtures thereof, did not support long term maintenance. Serum was required at a concentration of 5%, independent of other additives. Decreasing Ca++ levels below that normally found the basal medium (∼1 × 10−3) to as low as 1 × 10−4, resulted in increased short term proliferation, but decreased long term maintenance by causing a loss of stratification of the urothelium. Even a slight increase in Ca++ concentration from 1.0 to 1.5 × 10−3 resulted in an inhibition of proliferation and an increase in the number of large flat cells which subsequently sloughed off in sheets. The deletion of either insulin, hydrocortisone or both, inhibited growth. The addition of epidermal growth factor (EGF) or its homologue, transforming growth factor (TGF‐α), increased cell proliferation markedly and caused a variable increase in stratification. However, epithelium induced to rapid growth and proliferation with EGF, eventually exhausted its growth potential and died. TGF‐β1, alone or in combination with either EGF or α‐TGF, had no additional effect upon urothelial growth. Repeated transfers of urothelium by enzymatic dissociation led to decreased growth and maintenance potential. The data indicates that long term maintenance of stratified urothelium in culture requires a porous collagen substrate and fetal bovine serum together with hormonal requirements and concentrations of Ca++ that neither greatly stimulate nor inhibit growth.

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contrasting

confidence: 59%

Long term growth and maintenance of stratified rat urothelium in vitro

1989

Cell Proliferation

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“…The proteolytic enzyme trypsin can be used to separate bladder epithelium for in vitro studies (Bonar et al, 1977). Should trypsin have a similar action in vivo, then it might be possible selectively to remove malignant cells.…”

mentioning

confidence: 99%

Experimental Trypsin-induced Urothelial Cell Separationin vivo

Gardiner¹,

Masters

Molland³

1980

British Journal of Urology

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A morphological study of the effects of different concentrations of trypsin on the normal rat bladder in vivo is described. A low concentration (5000 u/ml) of the pure enzyme caused extensive urothelial cell separation following intravesical instillation via a catheter for 30 min. Urothelial regeneration commenced within 12 h and the appearance of the bladder was normal within 3 months. However, the development of submucosal haemorrhage and ulceration, partly as a result of infection, indicates that further experimental work is required before trypsin can be evaluated clinically for removing in situ carcinoma or reducing tumour bulk. The enzyme might also be used to produce more cellular material for cytological examination.

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“…2 and 3). The proteolytic enzyme, trypsin, can be used to separate bladder epithelium for in vitro studies (2,20), and Gardiner et al (9) reported the trypsin-induced urothelial cell separation in and observation of urothelial regeneration. The trypsinization which does not cause the disaggregation of cell layers lower than intermediate cells, was employed to remove superficial cells from normal mouse bladder epithelium (Figs.…”

Section: Discussionmentioning

confidence: 99%

Establishment of a Mouse Model of Cystitis and Roles of Type 1 Fimbriated Escherichia coli in Its Pathogenesis

Liao

Tomochika

Watanabe

et al. 1992

Microbiology and Immunology

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The role of type 1 fimbriae in promoting bladder colonization and the course of Escherichia coli cystitis were examined with type 1 fimbriated strains of clinically isolated E. coli. In the experiments of mice in vivo, intact bladder epithelium showed natural resistance to the adherence of type 1 fimbriated and nonfimbriated E. coli. However, the exfoliation of bladder superficial cells by trypsinization before the bacterial inoculation promoted the adhesion and colonization of type 1 fimbriated E. coli onto bladder epithelium. After colonization of E. coli, maximum numbers of E. coli and leukocytes were observed 3 days after inoculation.Nine days after inoculation, both of E. coli and leukocytes disappeared and the regeneration of superficial cells was observed.On the other hand, superficial cells in mice injected with phosphate-buffered saline or non-fimbriated E. coli regenerated 5 days after trypsinization.The present study demonstrated that the removal of superficial cells is essential for the adhesion and colonization of type 1 fimbriated E. coli onto bladder epithelium in vivo and a new model of E. coli cystitis in mice was established.The model which we established is valuable for histopathological, immunological, and therapeutic studies.

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Canine urinary bladder epithelial cells: Preparation for cell culture by enzyme dispersion

Cited by 16 publications

References 18 publications

Long term growth and maintenance of stratified rat urothelium in vitro

Long term growth and maintenance of stratified rat urothelium in vitro

Experimental Trypsin-induced Urothelial Cell Separationin vivo

Establishment of a Mouse Model of Cystitis and Roles of Type 1 Fimbriated Escherichia coli in Its Pathogenesis

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