A chemically defined medium for the propagation of rat prostate adenocarcinoma cells

Chan, Shamyuen

doi:10.1002/pros.2990020307

Cited by 4 publications

(4 citation statements)

References 13 publications

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“…Cell lines are therefore a valuable tool for the study of a variety of tumor biology questions related to prostatic cancer. Indeed, Shain et al [31] and Chen [32] have previously developed a series of useful rat prostatic cancer in vitro cell lines. In addition, Sestili et al [33] have established a cell line from the Dunning R3327-H tumor.…”

Section: Discussionmentioning

confidence: 99%

Establishment and characterization of seven dunning rat prostatic cancer cell lines and their use in developing methods for predicting metastatic abilities of prostatic cancers

et al. 1986

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In vitro cell lines were established from seven biologically distinct in vivo Dunning R3327 rat prostatic tumor sublines. Some of these in vitro cell lines (i.e., G, AT-1, AT-2) retain a low metastatic ability when inoculated back into syngeneic Copenhagen male rats, while others (i.e., AT-3, MAT-LyLu, MAT-Lu) retain a very high metastatic ability. A series of genetic (i.e., DNA content per cell, modal chromosomal number), as well as phenotypic parameters (i.e., morphology, 5 alpha-reductase, androgen receptor, estrogen receptor) were used to validate that the in vitro cell lines retained the major characteristics of the parental in vivo tumor sublines used for their respective establishment. A series of additional characteristics (i.e., morphology, growth rate, saturation density in surface culture, anchorage-dependent and -independent clonogenic potential) were compared between the high vs. the low metastatic in vitro cell lines to determine if a discriminatory parameter could be identified which reproducibly predicted the metastatic abilities of the particular prostatic cancer cell line. While the combination of the in vitro cell lines and their parental in vivo tumor subline will be a valuable tool for developing methods for predicting metastatic ability of prostate cancers, no single parameter yet measured is entirely successful in making this important distinction.

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Section: Discussionmentioning

confidence: 99%

Establishment and characterization of seven dunning rat prostatic cancer cell lines and their use in developing methods for predicting metastatic abilities of prostatic cancers

et al. 1986

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“…The rat prostate adenocarcinoma cell line PA-III was a cloned epithelial cell line which was originally derived from the prostate adenocarcinoma of an aged germ-free Lobund Wistar rat (Pollard, 1973;Chang & Pollard, 1977). The PA-III cells have been adapted to grow in a serum-free, chemically defined medium (Chan, 1981). The cells were propagated in the MB-DF medium plus penicillin (100 units/mL) and strep-©…”

Section: Methodsmentioning

confidence: 99%

Purification and properties of a plasminogen activator from cultured rat prostate adenocarcinoma cells

Strickland¹,

Serrano²,

Chan³

et al. 1983

Biochemistry

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Zymographic analysis of the supernates from confluent cultures of a rat prostate adenocarcinoma cell line, PA-III, revealed the existence of two molecular forms of specific plasminogen activators, one of molecular weight of approximately 80 000 and another of approximate molecular weight of 45 000, in sodium dodecyl sulfate. The low molecular weight form has been purified 364-fold in 66% yield from the culture medium by a combination of gel filtration on Sephacryl S-200 and affinity chromatography on Sepharose 4B-benzamidine. The purified material possessed a specific activity of 192 000 urokinase CTA units mg-1. This enzyme displayed activity toward human Glu1-plasminogen, characterized by a Km of 1.7 +/- 0.2 microM and a Vmax of 0.53 +/- 0.1 pmol of plasmin min-1 unit-1. A synthetic chromogenic substrate, H-D-Ile-Pro-Arg-p-nitroanilide (S-2288), was found for the activator. The enzyme possessed a Km of 0.33 mM and a kcat of 55 s-1 for S-2288. The activator was found to be a serine protease, inhibited by diisopropyl fluorophosphate (iPr2PF). At a concentration of 1 mM iPr2PF, and 30 nM enzyme, the half-time of this inhibition was 3.8 min. The 45 000 molecular weight enzyme was found to be inhibited by rabbit antibodies to human urokinase, thus characterizing the activator as a member of the urokinase class. The 80 000 molecular weight enzyme was not neutralized by anti-human urokinase but was neutralized by rabbit anti-human melanoma activator, likely allowing it to be classified as the tissue activator type.

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“…cell lines (PA-Ill and PA-IIIf) (Chang & Pollard, 1977;Chan, 1981) were used as target cells to investigate the oncolytic activity of VLDL. Both lines have been cloned and reconstitute adenocarcinomas when inoculated into LoW rats, where they metastasize spontaneously via ipsilateral lymphatic channels to the lungs and produce lesions (Pollard & Luckert, 1979;Chan, 1981). The PA-III cells were propagated in MEM with 10% heat-inactivated foetal bovine serum (Grand Island Biological Co., N.Y.), plus penicillin 100 u/ml) and streptomycin (100 [kg/ml).…”

Section: Methodsmentioning

confidence: 99%

“…The PA-III cells were propagated in MEM with 10% heat-inactivated foetal bovine serum (Grand Island Biological Co., N.Y.), plus penicillin 100 u/ml) and streptomycin (100 [kg/ml). The PA-IIJr line is a subclone of PA-Ill, which has been adapted to grow in a serum-free medium supplemented with growth factors (Chan, 1981). Colony-inhibition assay.-The colony-inhibiting assay was used to quantitate the cytotoxic activity of VLDL and its various subfractions.…”

Section: Methodsmentioning

confidence: 99%

Characterization of a very-low-density lipoprotein (VLDL)-associated cytotoxic factor

Chan¹,

Pollard²

1981

Br J Cancer

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Summary.-A VLDL-associated cytotoxic factor was isolated from sera of pregnant rats and characterized. The inhibitory effect of this factor on the macromolecular synthesis of rat prostate adenocarcinoma cells (PA-III) was also examined. VLDL (Sf 20-4C0) was subfractionated by differential ultracentrifugal flotation and the Sf 100-400 fraction was associated with most of the oncolytic activity. Chemical analysis of serum VLDL at various stages of pregnancy indicated that the 4 major constituents of VLDL (protein, triglyceride, cholesterol, and phospholipid), and the cytotoxic titre, were increased significantly before parturition and restored to normal levels by 24 h post partum. The delipidation of lyophilized VLDL by n-heptane suggested that the cytotoxic component was associated with the neutral lipid core of VLDL.Kinetic studies of colony inhibition and the incorporation of radioactive thymidine and leucine into 10% TCA precipitates of PA-III cells showed that VLDL induced irreparable cellular damage during the initial 15 h of incubation. The cytotoxic activity of VLDL was not due to the association with PGF2a,

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A chemically defined medium for the propagation of rat prostate adenocarcinoma cells

Cited by 4 publications

References 13 publications

Establishment and characterization of seven dunning rat prostatic cancer cell lines and their use in developing methods for predicting metastatic abilities of prostatic cancers

Establishment and characterization of seven dunning rat prostatic cancer cell lines and their use in developing methods for predicting metastatic abilities of prostatic cancers

Purification and properties of a plasminogen activator from cultured rat prostate adenocarcinoma cells

Characterization of a very-low-density lipoprotein (VLDL)-associated cytotoxic factor

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