Enzyme histochemistry of experimental embryo-derived teratocarcinomas

Damjanov, Ivan; Solter, Davor; Škreb, N.

doi:10.1007/bf00304029

Cited by 41 publications

(11 citation statements)

References 15 publications

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“…This would not be surprising, in view of the experimental origin of the teratoma from a 6-day postimplantation embryo (7). Moreover, the tumor stem cells have been found to resemble certain undifferentiated cells of the early postimplantation stage, rather than preimplantation-stage cells, in their ultrastructure (19) and alkaline phosphatase content (20); their soluble protein profiles also differ from those of morula cells (5). The evidence thus strongly supports the probability that some totipotent cells are normally still present as late as day 6 of embryonic life.…”

Section: Resultsmentioning

confidence: 99%

“…The evidence thus strongly supports the probability that some totipotent cells are normally still present as late as day 6 of embryonic life. The candidate totipotent cells have been tentatively identified as "ecto-mesoderm" (19,20), according to conventional "germ-layer" terminology. While germ layers normally arise in characteristic locations (for still unknown reasons) and normally undergo orderly movements ending in specific differentiations, experiments with many species (21), including mammalian embryos (22), have long shown that layers are substantially interconvertible and capable of giving rise to structures usually formed from the others.…”

Section: Resultsmentioning

confidence: 99%

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Totipotency and normal differentiation of single teratocarcinoma cells cloned by injection into blastocysts.

Illmensee¹,

Mintz²

1976

Proc. Natl. Acad. Sci. U.S.A.

398

169

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A definitive test for developmental totipotency of mouse malignant teratocarcinoma cells was conducted by cloning singly injected cells in genetically marked blastocysts. Totipotency was conclusively shown in an adult mosaic female whose tumor-strain cells had made substantial contributions to all of the wide range of its somatic tissues analyzed; the clonally propagated cell lineage had therefore differentiated in numerous normal directions. The test cells were from "cores" of embryoid bodies of a euploid, chromosomally male (X/Y), ascites tumor grown only in vivo by transplantation for 8 years. The capacity of cells from the same source to differentiate, in a phenotypic male, into reproductively functional sperms, has been shown in our previous experiments [(1975) Proc. Nat. Acad. Sci. USA 72, 3585-3589]. Cells from this transplant line therefore provide material suitable for projected somatic and germ-line genetic analyses of mammalian differentiation based on "cycling" of mutation-carrying tumor cells through developing embryos. In some animals obtained from single-cell injections tumor-derived cells were sporadically distributed in developmentally unrelated tissues. These cases can be accounted for by delayed and haphazard cellular integration, and by a marked degree of sustained cellular developmental flexibility in early mammalian development, irrespective of certain classical "germ-layer" designations. All mosaic mice obtained have thus far been free of teratomas. In one case, the injected stem cell contributed only to the pancreas and gave rise to a malignancy resembling pancreatic adenocarcinoma. The high modal frequency of euploidy in these individually tested cells thus tends to indicate that a near-normal chromosome complement is sufficient for total restoration of orderly gene expression in a normal embryonic environment; it may also be necessary for teratoma stem-cell proliferation to be terminated there.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Totipotency and normal differentiation of single teratocarcinoma cells cloned by injection into blastocysts.

Illmensee¹,

Mintz²

1976

Proc. Natl. Acad. Sci. U.S.A.

398

169

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“…Alkaline phosphatase was assayed by the method of Bessey et al (8), which follows the cleavage of p-nitrophenyl phosphate to p-nitrophenol by absorbance at 405 nm. One unit of enzyme activity is defined as 1 Amol of product per min at 370C.…”

Section: Methodsmentioning

confidence: 99%

“…Alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] has been the subject of a number of studies as a biochemical marker for the totipotential stem cells of the mouse teratoma. Damjanov et al (1) first cited the enzyme as a histochemical marker for these cells. Bernstine et al (2) showed that the proportion of stem cells in cell lines derived from the mouse teratoma cell line OTT-6050 could be correlated with alkaline phosphatase specific activity.…”

mentioning

confidence: 99%

Alkaline phosphatase of mouse teratoma stem cells: Immunochemical and structural evidence for its identity as a somatic gene product

Hass¹,

Wada²,

Herman³

et al. 1979

Proc. Natl. Acad. Sci. U.S.A.

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The immunochemical and structural characteristics of the alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] from mouse teratoma stem cells derived from the teratoma (ascitic and solid tumors and the F9 and PCC4 cell lines) have been compared to those of the alkaline phosphatases expressed in normal mouse placenta and several adult organs. Crossreactivity of the stem celalkaline phosphatase with antisera reacting with placental, kidney, liver, and brain alkaline phosphatases indicated that the stem cell enzyme had common antigenic determinants. Structural studies utilizing two-dimensional electrophoresis of the 32P-labeled alkaline phosphatase subunits showed that the stem cell, placental, and kidney alkaline phosphatases differed only in their sialic acid content and comigrated after removal of terminal sialic acid by neuraminidase digestion. Furthermore, one-dimensional peptide mapping of partial proteolysis fragments from 32P-labeled enzymes demonstrated identical fragmentation patterns for the stem cell and somatic enzymes. These immunochemical and structural data indicate that the stem cell alkaline phosphatase is the same core enzyme as that produced in the mouse placenta and kidney, with different amounts of terminal sialic acid. The one mouse alkaline phosphatase examined that differed from the other enzymes was the intestinal alkaline phosphatase. This isoenz me was not immunochemically crossreactive with the other arkaline phosphatases, did not comigrate in two-dimensional electrophoresis after neuraminidase digestion, and did not give identical peptide maps after partial proteolysis. Alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] has been the subject of a number of studies as a biochemical marker for the totipotential stem cells of the mouse teratoma. Damjanov et al.(1) first cited the enzyme as a histochemical marker for these cells. Bernstine et al. (2) showed that the proportion of stem cells in cell lines derived from the mouse teratoma cell line OTT-6050 could be correlated with alkaline phosphatase specific activity. The use of the stem cell alkaline phosphatase as a specific marker for the totipotential state of the stem cell would be dependent on the uniqueness of this gene product to the stem cell population. Based on the behavior of the enzymes in thermal inactivation and D-phenylalanine inhibition studies by Bernstine et al. (2), the stem cell enzyme was found to resemble the isoenzymes of mouse kidney and placenta but differed from the isoenzymes of mouse liver and intestine. Because functional studies on crude cell extracts are not adequate as a sole criterion for determining differences between gene products, Wada et al. (3) compared the enzymes of the ascitic and solid tumor forms of OTT-6050 with those of mouse kidney and placenta by using sodium dodecyl sulfate (NaDodSO4)/polyacrylamide gel electrophoresis and found that solid tumor and ascitic tumor enzymes had a subunit molecular...

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“…Several conclusions can be drawn from the RrdUrd-mith- [6][7][8] days. Also, the electrical rf-impedance properties of the BrdUrd-treated cells more closely resembled those of differentiated teratocarcinoma cells rather than the undifferentiated E C cells.…”

Section: Discussionmentioning

confidence: 99%

Flow cytometric analysis of the effect of 5‐bromodeoxyuridine on mouse teratocarcinoma cells

1980

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Several flow cytometric techniques were used to study the effects of 5-bromodeoxyuridine (BrdUrd) on the growth and differentiation of mouse teratocarcinoma cells. The teratocarcinoma system mimics embryogenesis, and the stem cells of this tumor, termed embryonal carcinoma (EC) cells, spontaneously differentiate in vitro to progeny that are morphologically, biochemically and biologically distinct from the EC cells. When cultures of EC cells were treated with 30 pM BrdUrd, a pleiotropic response was observed; i.e., certain parameters mimicked spontaneous differentiation but occurred 2-3 times more rapidly while others were inconsistent with, or diametric to, spontaneous differentiation. In one line of diploid EC cells in which tetraploidy accompanies spontaneous differentiation, the BrdUrd accelerated the appearance of the tetraploid population. Analysis of cell cycle distributions during the course of the BrdUrd treat--Murine testicular teratocarcinomas provide a suitable model for both mammalian development (8,11, 17) and differentiation in neoplasia (21 j. Embryonal carcinoma (EC) cells are the malignant stem cells of these tumors and are capable of differentiation in i i i w and in ritro to a variety of cell and tissue types, many of which are benign (15, 21).

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Enzyme histochemistry of experimental embryo-derived teratocarcinomas

Cited by 41 publications

References 15 publications

Totipotency and normal differentiation of single teratocarcinoma cells cloned by injection into blastocysts.

Totipotency and normal differentiation of single teratocarcinoma cells cloned by injection into blastocysts.

Alkaline phosphatase of mouse teratoma stem cells: Immunochemical and structural evidence for its identity as a somatic gene product

Flow cytometric analysis of the effect of 5‐bromodeoxyuridine on mouse teratocarcinoma cells

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