Gertrude Mungyer

Cell of the mucous membrane of the female genital tract in culture: A comparative study with regard to the histogenesis of endometriosis

SummaryCellular elements from the mucous membrane of the uterus and oviducts and from peritoneal washings were cultured. The in vitro behavior of these cells was compared to elucidate the histogenesis of endometriosis and the role of various diagnostic procedures.In 65% of the cultured matrial obtained by uterine-tubal flushing, proliferating cells of the uterine-tubal mucous membrane were present. Their morphology and behavior corresponded to those of cultured cells obtained by separate washing of the uterine cavity and the tubes, respectively, curetted material, and biopsies of endometriosis lesions.Epithelial and stromal cells were identified using phase contrast microscopy, electron microscopy, and immunohistochemical methods. These cell types did not occur in peritoneal washings before the flushing of uterus and tubes. It was therefore assumed that they were detached and transported to the pelvic cavity during the above-mentioned procedures. In view of their intensive proliferation they may form the basis in the development of nodules of endometriosis. This would support the implantation theory concerning the pathogenesis of endometriosis. Interactions between epithelial and mesothelial cells point to the possible role of the latter in encapsulating the endometrial elements.

Characterization of a hormone-producing ovarian carcinoma cell line

An ovarian carcinoma cell line (OTN 11) was produced from the ascitic fluid of a patient with a moderately to well differentiated papilliferous cystadenocarcinoma of the ovary. The cell line was characterized using electron microscopy karyotyping, immunohistochemical techniques with monoclonal antibodies against keratins as epithelial markers, and the monoclonal antibodies OV-TL 3 and OC 125 as ovarian carcinoma markers. These techniques revealed the epithelial and adenocarcinomatous nature of the cell line and the presence of ovarian carcinoma-related surface markers. The adenocarcinomatous nature of the cell line also became apparent after heterotransplantation of cell suspensions into nude mice and nude rats, in which adenomatous tumor structures were formed. These xenografts had the same ultrastructural and immunohistochemical properties as the cell line. Despite the adenocarcinomatous character of the tumor the cultured cells release estradiol into the culture medium. We may conclude that OTN 11 is an ovarian carcinoma cell line which has retained highly differentiated functions, such as the production of an ovarian hormone.

Microfilament assembly during lens cell elongation in vitro

Bovine epitheloid lens cells can be kept in culture for almost one year. During subculturing a gradual process of cell elongation is observed. Using several techniques, including immunofluorescence with antibodies against actin, electron microscopy, two-dimensional gel electrophoresis, drug treatment and measurement of actin polymerization by DNase I inhibition, it is shown that microfilament assembly parallels the process of cell elongation.

Changing intermediate-sized filament patterns in metastatic hepatocellular carcinoma cells of the guinea pig.

SummaryHepatocellular carcinoma cells obtained from ascitic fluid after diethylnitrosamine treatment of Sewall Wright strain-2 guinea pigs produce solid (primary) tumors, lymph-node metastases and malignant ascites when reinjected into animals of the same strain. When brought into culture the cells settle, form multilayer cultures and can be maintained in passage. In addition to epithelium-specific cytokeratin intermediate filaments (IF), these latter cells, like most cultured cells, also contain vimentin.Hepatocellular carcinoma cells in solid tumors and in metastatic tumors retain their original keratin IF and in general do not have an additional vimentin-IF system. When the tumor cells are present in ascites they develop vimentin-IF in addition to cytokeratin filaments. Vimentin is gradually lost when these cells sediment onto the peritoneal surface and proliferate continuously to form papillary projections, or when they are detected as circumscribed metastases. It seems likely, therefore, that in this system the synthesis of an additional vimentin cytoskeleton is related to reduced cell-to-cell contact and to the ability of the cells to survive individually or as cell clusters in body fluids, without being part of a cohesive tissue.

Plasmodium berghei: influence on granulopoiesis and macrophage production in balb/c mice.

Granulocyte and macrophage progenitor cells forming colonies in vitro (GM-CFC) from bone marrow, spleen, and peripheral blood of BALB/c mice infected with Plasmodium berghei were cultured at various times postinfection in a viscous, 0.8% methylcellulose system. The numbers of GM-CFCs from bone marrow increased gradually during the first week of infection, reaching a maximum around the tenth day of the disease. Subsequently, a rise of GM-CFCs in cultures of nucleated cells from the peripheral blood was observed and, with some delay, in spleen cell cultures also, with a maximum around the end of the second week. After the tenth day of malaria infection a fall of colony frequency in bone marrow-derived cells took place, leading to subnormal values of GM-CFCs during the third week of infection. Subsequently, a decrease in the spleen cell cultures followed, but colony numbers did not fall to normal values. The general increase in GM-CFCs in the different organs was preceded by a rise in serum levels of colony-stimulating activity (CSA), attaining a maximum 1 week after P. berghei inoculation. During the following period the CSA levels fell and reached normal values around the seventeenth day of the disease. Chemotherapy with chloroquine started on the fifteenth day of infection, when GM-CFCs in the bone marrow have dropped to normal values, stopped their further decrease. In the spleen a gradual normalization took more than 2 weeks. A challenge infection evoked an elevation of GM-CFC numbers in the bone marrow and in the spleen during the first 10 days in only about 50% of immune mice. The reaction was immediate in some animals, but generally lower and of shorter duration than during primary infection. The results have indicated that a lethal P. berghei infection in mice caused a transient increase in production of CSA followed by a general recruitment of GM-CFCs in all hemopoietic organs.

Changing tumour antigen expression in metastatic hepatocellular carcinoma cells of the guinea pig.

SummaryHepatocellular carcinoma cells (Line-10), obtained from ascitic fluid after diethylnitrosamine treatment of Sewall Wright strain-2 guinea pigs, produce solid (primary) tumours, lymphnode and lung metastases and malignant ascites when reinjected into animals of the same strain. Monoclonal antibodies were raised against the tumour cells by immunizing BALB/c mice with viable ascitic hepatocellular Line-10 tumour cells. Three hybridomas producing anti-Line-10 monoclonal antibodies were selected for further studies (10TL1, 10TL40 and 10TL43) and compared with monoclonal antibodies against intermediate filament keratins. The anti-Line-10 monoclonal antibodies did not cross react with Line-1 hepatocellular carcinoma cells, nor with normal guinea pig hepatocytes. When ascitic Line-10 cells form high papillary projections on the peritoneal surface, they significantly reduced their antigen expression of 10TL40 and of 10TL43 defined antigens, while the expression of 10TL1 defined antigens remained unaltered. Invading Line-10 cells in the deep submesothelial stromal tissue, however, lost reactivity with MoAb 10TL43 but not with the MoAb’s 10TL40 and 10TL1. The antigens on lung- and lymphnode metastases remained largely unaffected. The reactivity with MoAb’s 10TL40 and with 10TL43 was also lost upon prolonged culturing of Line-10 cells. The reactivity of Line-10 and Line-1 cells with all monoclonal antibodies against keratin filaments remained unaltered. Line-1 cells could be distinguished from Line-10 cells by the absence of any reactivity with the MoAb’s 10TL1, −40, −43, but also by the fact that 100% of the Line-1 cells were positive with antibodies against keratins 5/8, 18, and 7. The majority (>90%) of Line-10 cells, however, did not express the keratin 7. Since keratin 7 was also absent from guinea pig hepatocytes but strongly present on liver bile ductules it is suggested that the different antigenic make up of Line-1 and Line-10 cells might be related to a distinct cellular origin within the liver.