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Biomedical subjects

K R Stone

Publications and source records attributed to K R Stone.

12 recordsLinked to original sources

Isolation of a human prostate carcinoma cell line (DU 145).

A long-term tissue culture cell line has been derived from a human prostate adenocarcinoma metastatic to the brain. The cell line, DU 145, has been passaged 90 times in vitro over a period of 2 years. The cells are epithelial, grow in isolated islands on plastic Petri dishes, and form colonies in soft agar suspension culture. Karyotypic analysis demonstrates an aneuploid human karyotype with a modal chromosome number of 64. Distinctive marker chromosomes (a translocation Y chromosome, metacentric minute chromosomes and three large acrocentic chromosomes) have been identified. Electron microscopy of the original tumor tissue and of the tissue culture cell line show a remarkable similarity in cell organelle structure.

Acid Phosphatase

Investigation of human urogenital tract tumors of papovavirus etiology: brief communication.

Cells cultured from human urogenital cancer and other cancers as well as cells from noncancerous tissues were examined by immunofluorescent staining with antibodies to T-antigens and capsid antigens of papovaviruses BK virus (BKV), JC virus, and simian virus 40(SV40), and to capsid antigens of herpes simplex virus types 1 and 2 and human cytomegalovirus (CMV). Cells from early passage cultures of 123 primary tissues and from 14 continuous lines derived from transitional or renal cell carcinoma were tested. None of the cell preparations was specifically stained with any of the antisera. A serologic comparison of patients with bladder cancer, patients with prostate cancer, and normal control groups of BKV hemagglutination-inhibiting and SV40-neutralizing antibodies showed no differences among the 3 groups. None of the sera in the 3 groups had SV40 or BKV T-antibodies. In tests of supernatants of 35 primary cultures for presence of virus, a single isolation, that of a cytomegalovirus, was made. The study revealed no evidence that infection with papovaviruses of the SV40-polyoma subgroup has any part in the production of bladder and prostate cancer.

Animals

Scanning and transmission electron microscopy of human prostatic acinar cells.

Benign hyperplastic and neoplastic human prostate tissue samples were obtained by needle biopsy, transurethral resection or open prostatectomy. Acinar cells of both types of tissues were examined in the scanning electron microscope. It had been reported previously that adenocarcinoma acinar cells were more heterogeneous in size and shape than BPH acinar cells; the purpose of this study was to determine if there were surface morphology differences between the two types of tissues. Acinar cells were found to be extremely heterogeneous in their surface morphologies; three major types of surface morphologies were present - microvillous, ruffled, and bare. Within each class of surface morphology there was heterogeneity, both in size and density, of surface structures present. Microvillous, ruffled, and bare cells appeared to be present in normal, BPH, and neoplastic acini with no significant qualitative or quantitative differences in surface morphologies. Infrequently, it was possible to distinguish between well-differentiated and poorly-differentiated carcinomas because cells of the latter tissues were present in sheets rather than acini and appeared flat and totally devoid of surface detail. The SEM studies also sought to determine a marker to establish the origin of prostate tissue culture cells as normal, BPH or cancerous. Surface morphologies from tissues could be traced into the tissue cultures; again, three types of cells are present - bare, microvillous, and ruffled. However, since surface morphology does not appear to be a distinguishin feature of the pathology of the tissue it cannot provide a distinguishing marker for the origin of tissue culture cells. Scanning electron microscopy also provided an opportunity to observe possible secretory mechanisms and products in the prostate acinar cells.

Adenocarcinoma

Correlation of apparent molecular weight and antigenicity of viral proteins: an SDS-page separation followed by acrylamide-agarose electrophoresis and immunoprecipitation.

A simple method is described which combines a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SOS-PAGE) in the first demension with a second electrophoresis, at right angles to the first, into an agarose matrix. The proteins, separated by SDS-PAGE, are exposed to appropriate antisera after the second stage electrophoresis and immunoprecipitates form in the agarose corresponding to the relative electrophoretic mobilities of proteins in the first stage SDS-PAGE separation. The method thus provides a simple, reproducible means for correlating antigenicity with apparent molecular weight of proteins. The technique is qualtitative, but requires smaller quantities of antisera than more conventional immunoelectrophoretic methods such as rocket electrophoresis.

Antigens, Viral

Development and application of basic research techniques in bladder cancer research.

The growth of transitional epithelial cells with different growth media and growth supports was examined. Sephadex G-10, Bio-Gel P-20, Bio-Glas-1000, DEAE-Sephadex A-50, DEAE-cellulose, CM-Sephadex C-50, acid-soluble collagen, and immobilized collagen fibers were used to enhance plating efficiency. Acid-soluble collagen layers optimally increased the plating efficiency of primary cultures of bladder carcinoma. Media alterations with serial combinations of fetal calf, newborn calf, calf, bovine, and bull serum with minimum essential medium, Roswell Park Memorial Institute Tissue Culture Medium 1640, Connaught Medical Research Laboratories Medium 1066, Medium 199, Grand Island Biological, National Cancer Tissue Culture 135, 1415, McCoy's 5A, and National Cancer Institute medium were established. No promotion of cell division was noted with any one of these basic medium formulations.

Animals

Heterotransplantation of a human prostatic adenocarcinoma cell line in nude mice.

Nude mice of NIH/Swiss background were utilized for the heterotransplantation of a tissue culture cell line derived from a human prostate adenocarcinoma metastatic to the brain. These cells, which had been grown in vitro for 13 passages, formed solid tumors when injected s.c. into nude mice. The cell line DU 145 has been passaged 60 times in vitro over a period of 18 months. Tumors removed from the mice were serially transplanted to additional mice and reestablished in vitro. Light-microscopic analysis of the tumor grown in nude mice revealed a strong similarity to the patient's metastatic tumor. The ultrastructure of the tumor cells propagated in nude mice was compared to that of the original human tumor cells and to the tissue culture cells, both before and after passage in nude mouse. No major differences were detected. Karyotypic analysis of the tumor cells grown in vitro before mouse passage, grown in nude mouse, and grown in vitro after mouse passage indicated chromosomal identity and consistent marker chromosomes: three large acrocentric chromosomes and metacentric minute chromosomes.

Adenocarcinoma

Scanning electron microscopy of hyperplastic and neoplastic human prostate.

Acinar cells of neoplastic prostate tissues are more heterogeneous in size and shape than benign hyperplastic cells when observed by scanning electron microscopy. Three types of acinar cells are recognizable by surface structure, cells with microvilli, cells without microvilli, and cells with membrane ruffles. The pitted cells previously seen in BPH tissues are probably artifactual. The identity of the crater cells is sitll in question.

Adenocarcinoma

In vitro cultivation of prostatic epithelium.

A tissue culture procedure for in vitro cultivation of human prostatic tissue that utilizes the enzyme collagenase to disrupt the prostate specimen is described. Collagenase effectively disperses the tissue stroma into single cells which are easily separated from the largely intact glandular structures. The isolated glandular fragments then are used to initiate monolayers of prostatic epithelium in vitro.

Cell Separation

Morphologic and immunologic studies of human prostatic carcinoma.

Benign and malignant prostatic tissue was removed in surgery and partitioned for (a) ultrastructural study, (b) tissue culture, and (c) (c) immunochemical study. Fourteen malignant and 18 benign prostatic cancer specimens were examined by transmission electron microscopy (TEM) for the presence of viruses or virus-like particles. Viruses could not be identified with assurity in thin sections. Acinar cells of normal, benign prostatic hypertrophy (BPH), and neoplastic prostate tissue were examined in the scanning electron microscope and TEM and found to be extremely heterogenous in their surface morphologies. Three major types of surface morphologies were present: microvillous, ruffled, and bare. All three types of cells were present in normal, BPH, and neoplastic acini. A collagenase procedure was utilized to remove the stromal cells from glandular structures prior to in vitro cultivation. Partially purified extracts from 71 human urothelial tumors and 75 human urothelial nontumor tissues were used as competing antigens in competition radioimmunoassay in an effort to detect the presence of one of the structural components of type-C ribonucleic acid viruses, the p30 core protein. The urothelial tumors tested included 42 BPH specimens and 18 prostatic carcinoma specimens. Thirty-eight percent of the prostatic carcinoma tissues and 48% of the BPH tissues demonstrated the presence of a protein antigenically similar to the p30 core protein of an oncogenic RNA virus.

Adenocarcinoma