Hormonal induction of differentiation in teratocarcinoma stem cells: Generation of parietal endoderm by retinoic acid and dibutyryl cAMP (original) (raw)
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The induction of antigenic changes in a teratocarcinoma stem cell line (F9) by retinoic acid
Developmental Biology, 1979
Treatment of embryonal carcinoma cells F9 with retinoic acid results in the appearance of epithelioid cells resembling endoderm which synthesize basement membrane protein and plasminogen activator. Concomitant with the appearance of these properties of differentiated cells, the epithelial cells cease to express SEA-l, an antigenic determinant characteristic of teratocarcinoma stem cells and early mouse embryos. Our evidence indicates that the phenotypic changes that accompany retinoic acid treatment of embryonal carcinoma cells are irreversible and a consequence of the differentiation of the cells into endoderm 515
Journal of Cellular Physiology, 1990
F9 teratocarcinoma stem cells differentiate into parietal endoderm-like cells when given retinoic acid (RA) and dibutyryl cyclic adenosine monophosphate (DB-cAMP). It is generally accepted that the stem cells are resistant to the action of cAMP alone and need to be primed by RA in order to respond to cAMP. In this report, we demonstrate that F9 stem cells differentiate into parietal endoderm-like cells in the absence of exogenous RA when treated with cholera toxin and 1-methyl,3-isobutyl xanthine (CT/MIX) or 8-bromo-cAMP/MIX (8B2-cAMP/MIX). Cells treated with CT/MIX or 8B2-cAMP/MIX were morphologically similar to parietal endoderm-like cells, produced high amounts of plasminogen activator, and synthesized both type IV collagen and laminin mRNA. Conversely, markers made in abundance by stem cells such as stage-specific embryonic antigen (SSEA-1) and an mRNA species of 6.8 kb (pST6-135) were markedly reduced in CT/MIX-treated cells. To prove that cAMP alone could induce differentiation Lipidex-1000, a hydrophobio gel, was used to remove 80–;90% of the endogenous serum retinoids. F9 cells grown in this retinoid-depleted serum and treated with 8B2-cAMP/MIX differentiated to parietal endoderm-like cells as shown by both dramatic changes in morphology and induction of type IV collagen mRNA. Our results indicate that the differentiation of F9 to parietal endoderm-like cells can be induced by increased intracellular cAMP and is not strictly dependent on the addition of RA.
Physiological Research
In both embryonal carcinoma (EC) and embryonic stem (ES) cells, the differentiation pathway entered after treatment with retinoic acid (RA) varies as it is based upon different conditions of culture. This study employs mouse EC cells P19 to investigate the effects of serum on RA-induced neural differentiation occurring in a simplified monolayer culture. Cell morphology and expression of lineage-specific molecular markers document that, while non-neural cell types arise after treatment with RA under serum-containing conditions, in chemically defined serum-free media RA induces massive neural differentiation in concentrations of 10(-9) M and higher. Moreover, not only neural (Mash-1) and neuroectodermal (Pax-6), but also endodermal (GATA-4, alpha-fetoprotein) genes are expressed at early stages of differentiation driven by RA under serum-free conditions. Furthermore, as determined by the luciferase reporter assay, the presence or absence of the serum does not affect the activity of th...
Cell Differentiation, 1987
Several subclones of the human embryonal carcinoma (EC) cell line Tera-2 can be induced to differentiate in monolayer culture by retinoic acid (RA) to a flattened cell type with reduced growth rate. Using a method based on the transition probability model, we have analysed changes in cell cycle kinetics of Tera-2 cells during the differentiation process. Growth inhibition was shown to occur without a lag period and to be partly due to an increase in the duration of the S-phase, but with a relatively greater contribution from an increase in the duration of Gl-phase. Since the fraction of the cell population in the Gl-phase then doubled, cells accumulated in this part of the cycle. In contrast, the reduced proliferation rate of two murine EC cell lines, PC13 and P19, treated with RA occurs after a lag period of about two cell cycles and is mainly attributable to an increase in the duration of the S-phase. The results illustrate a differential response of human and murine EC cells to growth regulation by RA and again emphasize that although the stem cells of murine teratocarcinomas may provide a useful model, they are not identical to their human counterparts. Cell cycle analysis; Human teratocarcinoma; Retinoic acid; Differentiation
Differentiation, 1999
The vimentin gene encodes an intermediate filament protein expressed in the parietal endoderm, mesodermal, and early neural cells in vivo but by most invitro-cultured cells regardless of their embryonic origin. Here we show that the vimentin gene promoter is very active in F9 embryonal carcinoma cells and increases in activity during differentiation. Using a series of 5′deletion mutants, we provide evidence that the regions of the promoter involved in F9 cell activity are different from those previously demonstrated to be active in differentiated cell lines. Furthermore, we show that in differentiating F9 cells the activities of two different regions of the promoter are significantly enhanced. A distal region (-1710/-957) appears to contain functional binding sites for the murine Hox-A5 homeoprotein as demonstrated by band shift and footprinting experiments. A proximal region (-140/-78) contains a 30-bp repetitive sequence found in other genes activated during differentiation of F9 cells. Using band shift assays and methylation interference, we present evidence that a sequence-specific single-stranded DNA-binding protein(s) specifically interacts with the minus strand of the 30-bp sequence.
Retinoic acid-induced neural differentiation of embryonal carcinoma cells
… and cellular biology, 1983
We have previously shown that the P19 line of embryonal carcinoma cells develops into neurons, astroglia, and fibroblasts after aggregation and exposure to retinoic acid. The neurons were initially identified by their morphology and by the presence of neurofilaments within their cytoplasm. We have more fully documented the neuronal nature of these cells by showing that their cell surfaces display tetanus toxin receptors, a neuronal cell marker, and that choline acetyltransferase and acetyl cholinesterase activities appear coordinately in neuroncontaining cultures. Several days before the appearance of neurons, there is a marked decrease in the amount of an embryonal carcinoma surface antigen, and at the same time there is a substantial decrease in the volumes of individual cells. Various retinoids were able to induce the development of neurons in cultures of aggregated P19 cells, but it did not appear that polyamine metabolism was involved in the effect. We have isolated a mutant clone which does not differentiate in the presence of any of the drugs which are normally effective in inducing differentiation of P19 cells. This mutant and others may help to elucidate the chain of events triggered by retinoic acid and other differentiation-inducing drugs.
Cancer research, 1988
We have examined the effects of dietary retinoids upon the growth and differentiation of seven embryonal carcinoma lines in mice. The control diet contained 4000 IU/mg retinyl palmitate; the other diets contained 2 x 10(5) IU/mg retinyl palmitate, 50 mg/kg all-trans-retinoic acid (RA), 100 mg/kg RA, and no retinoid. The RA-containing diets had little influence on tumor latency or incidence but did suppress growth of many of the tumors. Decreased tumor mass was, in most but not all instances, accompanied by an increased proportion of differentiated cells. Increased differentiation was most commonly quantitative rather than qualitative; i.e., there was a larger proportion of the same types of differentiated cells seen in tumors from the control diet group rather than an increase in the spectrum of cell types observed. Notably, tumors from two differentiation-defective embryonal carcinoma lines were refractory to both the differentiation-inducing and growth-suppressing properties of di...
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~.