Flow Cytoenzymology of the Early Differentiation of Mouse Embryonal Carcinoma Cells (original) (raw)
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Ultrastructural localization of membrane phosphatases in teratocarcinoma and early embryos
The American journal of pathology, 1977
Ectodermal cells of the two- and three-germ layer-thick mouse egg-cylinders are considered to be the progenitors of embryonal carcinoma cells in embryo-derived teratocarcinomas. In an attempt to find differences between the tumor cells and equivalent embryonic cells, we have studied the electron microscopic cytochemical localization of alkaline phosphatase, 5'-nucleotidase, and Mg2+-activated adenosine triphosphatase (ATPase) in embryo-derived teratocarcinomas and mouse egg-cylinders. Alkaline phosphatase was detected in both embryonic and tumor cells, but its activity appeared much more intense in the tumor cells. No ATPase was demonstrated in embryonic ectodermal cells of 6-day-old embryos and only in occasional cells of 7- and 8-day-old embryos. No 5'-nucleotidase activity could be demonstrated in 6- to 8-day-old cylinders. There was marked ATPase and 5'-nucleotidase activity in the membranes of embryonal carcinoma cells. These data point out some differences on the p...
Growth and differentiation of an embryonal carcinoma cell line (C145b)
Development, 1979
Several cell and tumour lines were isolated from a single-embryo-derived teratocarcinoma and their karyotypes and differentiation in adult hosts recorded. The majority of cells contained normal karyotypes by banding. The cells were injected into blastocysts and although they sometimes colonized the yolk sac, they never colonized the embryo. Thus the possession of a normal karyotype is not a sufficient condition for embryo colonization. The loss of growth capacity was investigated by studying differentiation and tumourigenicity in a variety of circumstances. The change in appearance from an EC cell morphology to a big flat cell in culture leads to retardation of growth in adult hosts. When EC cells are injected into a blastocyst, the ability to grow progressively both in culture and in adult hosts is lost.
Human embryonal carcinoma grown in athymic mice and in vitro
Cancer research, 1980
Tissue from 19 human testis tumors was transplanted into athymic mice. One embryonal carcinoma, ECCS, grew rapidly, and this tumor was studied both as a xenograft and an in vitro culture of xenograft-derived tumor cells. Xenografts showed no evidence of differentiation. The embryonal carcinoma cells were heteroploid and showed alkaline phosphatase activity. When tumor cells from the xenografts were grown in vitro, the cells formed aggregates resembling embryoid bodies with epithelium-like cells in the periphery. Regularly, another population of mouse cells which showed several criteria of malignancy overgrew the culture and could be subcultured continuously. These abnormal cells may result from an in vivo or in vitro transformation of mouse stromal cells.
Cancer Research, 1985
A431 human epidermoid carcinoma cells monophenotypically express the placenta! alkaline phosphatase (PLAP)-like enzyme shown by its catalytic and antigenic characteristics, properties which are shared by the Nagao isozyme. More specifically, it is L-leucine sensitive just as is the rare placenta! D-variant of PLAP and the testicular heat-stable enzyme. Collectively, these are all referred to as PLAP-like enzymes. The enzyme was localized to the surface of the plasma mem brane since it was released in an active form by bromelain treatment of cells. The number of molecules per A431 cell was estimated by radioimmunoassay at 7.5 x 105, a value signifi cantly higher than that observed for HeLa TCRC-1 cells (5 x 104) which express the S-variant of PLAP, also referred to as the Regan isozyme. The quantity of the enzyme was increased significantly (10-fold) by treating the cells with modulating agents including sodium butyrate, prednisolone, and hyperosmolar so dium chloride. The identification of a cell line such as A431 with enhanced expression in the amount of the PLAP-like enzyme and which can be further enhanced by modulating agents will facilitate studies of the differences and the similarities between this protein and other variants of PLAP. The A431 cell line now takes its place with other cell lines which are phenotypically restricted in their expression of alkaline phosphatase. Finally, the A431 cell line is also shown here to be a suitable model system for in vivo tumor studies ;>uch as immunolocalization.
Cancer research, 1973
The DNA:protein ratio and enzyme patterns of four spontaneous mouse tumors (one male and one female mammary carcinoma, a muscle sarcoma, and a uterine fibrosarcoma) were determined. Tissue culture cell lines established from these tumors and from mouse embryo and human embryo lung tissues showed quantitative changes in DNA:protein ratio and most of the 13 enzymes studied. Although there were wide variations in enzyme patterns between the tumors and between the embryo tissues, the pattern in tissue culture cells, whatever their origin, tended to be similar. Most of the tumor cell enzyme changes were reversible after reimplantation into syngeneic hosts, with the exception of alkaline phosphatase, which did not reappear in muscle sarcoma and female mammary carcinoma derived from tissue culture cells. In tissue culture, the mitochondrial enzymes succinic dehydrogenase and cytochrome oxidase were particularly affected. Surface enzymes were also altered. Alkaline phosphatase was decreased or lost and 5'-nucleotidase was increased in all cultures. The enzymes of the male mammary carcinoma were less affected by culture than those of the other tumors.
Factors influencing the differentiation of embryonal carcinoma and embryo-derived stem cells
Experimental Cell Research, 1989
The effects of aggregation, retinoic acid, and medium conditioned by Buffalo rat liver (BBL) cells, alone and in combination, on the differentiation of PSA4TG12 embryonal carcinoma and El4 embryonal stem cells are reported. The observations indicate that BBL-conditioned medium has more than one effect on the differentiation process, that retinoic acid has at least two effects which operate in different concentration ranges, and that both agents influence the choice of differentiation pathway as well as the extent of differentiation. @ 1989 Academic RUSS, 1~.
Alkaline phosphatase activity and the regulation of growth in transformed mammalian cells
Journal of Cellular Physiology, 1974
The relationship between alkaline phosphatase activity and cell growth has been studied in hamster cells transformed by different carcinogens. About 90% of normal hamster embryo cells were constitutively positive for alkaline phosphatase activity (AP+). However, there were no AP+ cells in cell lines transformed after treatment with the chemical carcinogens dimethylnitrosamine or 4‐nitro‐quinoline‐N‐oxide and 0.02% and 4% AP+ cells in cell lines transformed by polyoma virus or Simian virus 40. The glucocorticoid hormone, prednisolone, induced alkaline phosphatase activity in 12% and 44% of the enzyme‐negative (AP−) cells in cell lines transformed by polyoma or Simian virus 40, but this hormone did not induce alkaline phosphatase activity in AP− cells from cell lines transformed after treatment with the chemical carcinogens. Treatment of polyoma transformed AP− cells with the mutagen N‐methyl‐N′‐nitro‐N‐nitro‐soguanidine produced AP+ cells, whereas no AP+ cells were found after mutage...
British journal of cancer, 1994
Placental and placental-like alkaline phosphatase (PLAP) levels in the culture media of 87 cell lines of neoplastic and 'normal' origin were measured by a conventional immunosorbent enzymatic assay (IAEA) and by a new immunoradiometric assay (IRMA). The IRMA detected immunoreactive PLAP in 37 of 80 (46%) human epithelial and germ cell cultures, while the IAEA detected PLAP in only 25 (33%). Of the 52 non-germ cell tumour cultures, the IRMA detected expression in 24 (46%) and the IAEA in only 16 (31%). In 17 cases (21%) the IRMA recorded levels double that of the IAEA, while in five cultures (6%) the reverse was true. The IRMA was much more robust than the IAEA and had considerably lower inter- and intra-assay coefficients of variation (3.75-8.5% vs 5.2-46%). Detection of PLAP(-like) expression by IAEA is dependent on neoplastic expression of enzymatically functional molecules and quantification assumes constant enzyme kinetics. PLAP-like material has a higher catalytic rate ...
Ultrastructural differentiation of a clonal human embryonal carcinoma cell line in vitro
Cancer research, 1983
A cloned human embryonal carcinoma (EC) cell line 2102Ep derived from a testicular teratocarcinoma was characterized by means of electron microscopy and immunohistochemistry. These EC cells when plated at high cell density grow mostly as undifferentiated cells displayed relatively little pleomorphism. Eighty-five to 90% of these cells contain keratin in the form of peridesmosomal tonofilaments. Cell populations of the same clonal line plated at a low cell density contain, in addition to undifferentiated EC cells, large cells displaying complex cytoplasmic architecture, more complex junctions, and intracytoplasmic keratin in the form of bundles. Some of these cells also react with antibodies to human chorionic gonadotropin indicative of trophoblastic differentiation. Furthermore, some cells form "morules" which are multicellular aggregates composed of a core of EC cells and an attenuated, more differentiated outer cell layer. These data thus point out not only some similari...
Embryonal carcinoma cells are the undetermined stem cells of teratocarcinomas. Supplementation of culture medium with P-mercaptoethanol permits the feeder layer independent clonal growth and differentiation of normally feeder layer dependent embryonal carcinoma cell lines. Differentiated cells within the clones appeared less than 6 days after plating and were distinguished from embryonal carcinoma cells by their morphology, lack of histochemically detectable alkaline phosphatase activity, and secretion of plasminogen activator. Over 70% of the colonies secreted plasminogen activator after 6 days.