Isolation of differentially expressed human cDNA clones: similarities between mouse and human embryonal carcinoma cell differentiation

Development ◽  
1988 ◽  
Vol 104 (3) ◽  
pp. 403-413
Author(s):  
M.V. Wiles
Keyword(s):  
Cell Differentiation ◽  
Human Development ◽  
Cell Line ◽  
Cdna Library ◽  
Embryonal Carcinoma ◽  
Cdna Clones ◽  
Embryonal Carcinoma Cell ◽  
Visceral Endoderm ◽  
Induced Differentiation ◽  
Ec Cells

The study of early human development is of great importance but has been limited by the lack of suitable reagents. Recently, however, the human embryonal carcinoma (EC) cell line NT2D1 has been isolated. This cell line will differentiate upon exposure to retinoic acid (RA). A cDNA library was constructed from poly(A)+ RNA derived from NT2D1 cells treated with 10(−5) M-RA for 7 days (delta NT2D1 cells). By differential cDNA screening, it was found that 1.12% of delta NT2D1 cDNA recombinants screened detected an increase in signal with 32P-cDNAs derived from delta NT2D1 as compared with NT2D1. To compare RA-induced differentiation of mouse and human EC cells, the delta NT2D1 cDNA library was rescreened with 32P-cDNAs derived from the mouse EC cell line F9 and the result compared with 32P-cDNA derived from F9 differentiated to parietalendoderm (F9PE)-like cells and visceral-endoderm (F9VE)-like cells. Approximately 1.2% of the delta NT2D1 cDNA recombinants detected a differential increase in signal following differentiation of mouse EC cells to F9VE and/or F9PE. Of these homologous regulated sequences, 0.3% were common to both mouse and human EC cell RA-induced differentiation. Five different cDNA clones were isolated that detect a marked increase (5- to 75-fold) in mRNA abundance following RA-induced differentiation of NT2D1. Of these five clones, three detect homologous mRNAs which also increase in abundance following differentiation of the mouse EC cell line F9 to PE- and/or VE-like cells; the other two clones do not detect sequences in the mouse mRNAs tested. One clone shows homology to SPARC, a gene known to be regulated during mouse embryonic development. While another clone, SO5A, has a limited range of expression, being detected in F9VE and in a human parietal-endoderm-like cell, but not in F9PE and a human visceral-endoderm-like cell. This work shows that there are both similarities and differences in mouse and human EC cell differentiation, and these cDNA clones provide some of the first reagents for studying the molecular biology of human development.

scholarly journals States of developmental commitment of a mouse embryonal carcinoma cell line differentiating along a neural pathway.

1989 ◽  
Vol 109 (5) ◽  
pp. 2481-2493 ◽  
Author(s):  
E Lang ◽  
M L Mazauric-Stüker ◽  
A Maelicke
Keyword(s):  
Retinoic Acid ◽  
Cell Line ◽  
Embryonal Carcinoma ◽  
Carcinoma Cell ◽  
Neuronal Cell ◽  
Carcinoma Cell Line ◽  
Mammalian Brain ◽  
Embryonal Carcinoma Cell ◽  
Induced Differentiation ◽  
Embryonal Carcinoma Cell Line

The embryonal carcinoma cell line PCC7-S-AzaR1 (clone 1009) has been shown to differentiate in the presence of all-trans retinoic acid and dibutyryl cAMP into cells of predominantly neural properties (Paulin, D., H. Jakob, F. Jacob, K. Weber, and M. Osborn. 1982. Differentiation. 22:90-99). By analyzing the marker expression of derivatives in further detail, we characterized the two major cell phenotypes as neuron- and fibroblast-like and the two minor ones as astroglia- and endothelial-like. The stability of developmental commitment of clone 1009 was tested by recloning. The isolated subclones exhibited different patterns of chemically induced derivatives, with some of them (denoted N-clones) producing only a single (neuronal) cell type. As shown by long-term cultures in the absence of retinoic acid, the properties of isolated subclones remained essentially stable. In contrast to the clones producing neuron-like and other derivatives upon induced differentiation, the (exclusively neuronal) derivatives of N-clones detached and died within a few days in culture. If maintained in the presence of other neural cell types, however, their survival was dramatically extended indicating a requirement for specific interactions with other cells of the same tissue. The patterns of derivatives obtained from N-clones depended on the chemical nature of the substrate on which they were grown. Thus, when seeded on laminin-coated surfaces before induced differentiation, N-clones developed not only to neuron-like derivatives but rather to the same four derivatives observed with the original cell pool. These and further results suggest a common cell lineage of the identified phenotypes. The isolated subclones of uninduced cells probably represent different states of commitment within the same developmental pathway. Their stability offers the opportunity to analyze the nature of cellular commitment on the cellular, molecular, and genetic levels. This makes the family of clones derived from PCC7-S-AzaR1 (clone 1009) cells an advantageous in vitro model of mammalian brain early ontogenesis.

Growth and differentiation of an embryonal carcinoma cell line (C145b)

Development ◽  
1979 ◽  
Vol 54 (1) ◽  
pp. 277-295
Author(s):  
V. E. Papaioannou ◽  
E. P. Evans ◽  
R. L. Gardner ◽  
C. F. Graham
Keyword(s):  
Cell Line ◽  
Cell Morphology ◽  
Embryonal Carcinoma ◽  
Carcinoma Cell ◽  
Normal Karyotype ◽  
Embryonal Carcinoma Cell ◽  
Sufficient Condition ◽  
Embryonal Carcinoma Cell Line ◽  
Ec Cells ◽  
Flat Cell

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.

Cell cycle analysis during retinoic acid induced differentiation of a human embryonal carcinoma-derived cell line

1987 ◽  
Vol 20 (2-3) ◽  
pp. 153-160 ◽  
Author(s):  
Christine L. Mummery ◽  
Marga A. van Rooijen ◽  
Stieneke E. van den Brink ◽  
Siegfried W. de Laat
Keyword(s):  
Cell Cycle ◽  
Retinoic Acid ◽  
Cell Line ◽  
Embryonal Carcinoma ◽  
Cell Cycle Analysis ◽  
Induced Differentiation

The mode of cell death associated with cavitation in teratocarcinoma-derived embryoid bodies

Development ◽  
1984 ◽  
Vol 80 (1) ◽  
pp. 63-74
Author(s):  
Sheilagh M. Boyd ◽  
M. L. Hooper ◽  
A. H. Wyllie
Keyword(s):  
Cell Death ◽  
Cell Line ◽  
Embryonal Carcinoma ◽  
Carcinoma Cell ◽  
Embryoid Bodies ◽  
Embryonal Carcinoma Cell ◽  
Embryonal Carcinoma Cell Line ◽  
Related Cell ◽  
Significant Difference ◽  
Tissue Modelling

Cell death occurring in embryoid bodies derived from the embryonal carcinoma cell line, PSA4, which undergo cavitation, and in those from the related cell line S2, which do not undergo cavitation, was classified as apoptosis or necrosis by ultrastructural criteria. Both modes of cell death were seen in PSA4 embryoid bodies while apoptosis alone was seen in S2 embryoid bodies. No significant difference was found between PSA4 and S2 embryoid bodies either in apoptotic incidence score or in the spatial distribution of apoptotic events. We therefore conclude that although apoptosis and tissue modelling coexist in PSA4 embryoid bodies, necrosis rather than apoptosis is causally related to formation of the cavity.

Coordinate expression of parietal endodermal functions in hybrids of embryonal carcinoma and endodermal cells

1982 ◽  
Vol 2 (3) ◽  
pp. 331-337
Author(s):  
W E Howe ◽  
R G Oshima
Keyword(s):  
Cell Line ◽  
Embryonal Carcinoma ◽  
Hybrid Cell ◽  
Cytoskeletal Proteins ◽  
Characteristic Functions ◽  
Embryonal Carcinoma Cell ◽  
Coordinate Expression ◽  
Embryonal Carcinoma Cell Line ◽  
Endodermal Cells ◽  
Hybrid Clones

A derivative, FOT5, of the F9 murine embryonal carcinoma cell line which is resistant to ouabain and thioguanine was fused with a near diploid parietal endodermal cell line, PFHR9, Hybrid clones (ENEC1 to ENEC5) were isolated in HAT Medium containing ouabain at a frequency of approximately 2 x 10(-4). The DNA contents and chromosome number of the ENEC hybrids were approximately the sum of those of the parents. Five hybrid cell lines examined in detail expressed the following parietal endodermal functions: plasminogen activator activity, basement membrane proteins, and endodermal cytoskeletal proteins. Embryonal carcinoma characteristic functions (tumorigenicity, a stage specific embryonic antigen, and high alkaline phosphatase activity) were extinguished in the hybrids. No hybrid clones with embryonal carcinoma morphology were observed among 1,358 hybrid clones examined. Hybrids, propagated for over 100 generations, continued to express endodermal functions and not embryonal carcinoma functions. The coordinate expression of endodermal functions and the extinction of embryonal carcinoma functions in the ENEC hybrids suggest that the parietal endodermal cells contain diffusible activities which extinguish embryonal carcinoma functions and possibly cause the embryonal carcinoma genome to express parietal endodermal characteristics.

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