Teratocarcinoma cells

Two methods have been developed to establish mouse teratocarcinomas cells in culture. 

1) Tumour cells are allowed to attach to gelatin-coated tissue culture dishes (see 2.4.1) when many different kinds of cells including embryonal cells grow out. These latter cells may eventually outgrow the differentiated cells and they may then be cloned (Chapter 7) (Rosenthal et al., 1970 Bernstine et al., 1973). Lines isolated in this way tend after some time to lose their ability to differentiate. 

2) Tumour cells (ascites or dissociated solid tumours) are plated out on a monolayer of feeder fibroblasts (see 7.1.4). After several days many different sorts of differentiated cells may be seen and also nests of embryonal cells. On repeated subculture these cells 

Once isolated the undifferentiated cells remain homogeneous as long as they are subcultured using 0.25% trypsin, 0.05 mM EDTA in PBS (Chapter 4) into Dulbecco s modified Eagle s medium (Appendix 1) supplemented with 5-20% bovine serum before they become confluent. 

If the teratocarcinoma cells become confluent they will begin to differentiate into many different cells types, but this process is not so dramatic as the differentiation which occurs if cells isolated by method 2 are transferred to a vessel without a feeder layer. Then the cells attach very poorly and form clumps in suspension. These clumps remain healthy and quickly differentiate (Evans, 1972) to form an outer layer of endoderm cells. The presence of endoderm can be shown by assaying for the serine protease plasminogen activator which is a marker typical of endoderm cells (Strickland et al., 1976). 

---

Bradley, A., Evans, M., Kaufman, M.H., and Robertson, E. (1984) Formation of germ-line chimaeras from embryo derived teratocarcinoma cell lines. Nature 309, 255-256. 

A number of companies supply dishes and TC trays precoated with extracellular matrix collagen, or gelatin (e.g. International Biotechnologies Ltd., and Bibby Science Products Ltd.). This promotes attachment and flattening and is essential for growth of cells such as the F9 teratocarcinoma cells. 

Fig. 16.1 Both teratocarcinoma cells and normal embryonal stem cells behave alike. Both types of cells are accepted and incorporated into the inner layer of the blastocyst, when they are injected into the blastocyst cavity. They behave like normal embryonic stem cells, and the progeny of these cells are found in practically every cell and tissue of the chimeric mouse, where they differentiate normally. Moreover, they even form normal germ cells. Thus, teratocarcinoma stem cells, when separated from their undifferentiated cancerous daughter cells and transplanted into a compatible host, not only survive, they also do no harm to the host and do not make him cancerous. Incidentally, the capability of embryonic stem cells to be accepted by a recipient blastocyst is the basis of producing gene knock-out, chimeric mice, where a normal gene is replaced by an altered, engineered version of the gene. These chimeric mice then carry the mutation in their stem cells and transmit them to differentiated cells, where they are expressed. (With permission of Taylor and Francis, Inc. See Rg. 21-32 in ref. 1.)...

Kojima N, Fenderson B, Stroud M, Goldberg R, Habermann 36. R, Toyokuni T, Hakomori S. Further studies on cell adhesion based on Le -Le interaction, with new approaches embryo-glycan aggregation of F9 teratocarcinoma cells, and adhesion 37. of various tumour cells based on Le expression. Glycoconj. J. 1994 11 238-248. 

Rusciano, D., Lorenzoni, P. and Burger, M. M. (1992). Specific growth stimulation in the absence of specific cellular adhesion in lung colonization by retinoic acid treated F9 teratocarcinoma cells. Int. J. Cancer 52, 471-477. 

A pluripotent human teratocarcinoma cell clone, NT2/D1, which was derived from the Tera-2 cell line, was induced to differentiate into cells with neuronal cell morphology by treatment with berberine. As early as one day after a 24 hour treatment of cells with berberine at a non-toxic dose of 0.1 mg/ml in culture medium, the cells began to show morphologic... 

Hosier, B.A., Rogers, M.B., Kozak, C.A., and Gudas, L.J. (1993). An octamer motif contributes to the expression of the retinoic acid-regulated zing finger gene Rex-1 (Zfp-42) in F9 teratocarcinoma cells. Mol. Cell Biol. 13 2919-2928. 

Illmensee, K., and Mintz, B. (1976). Totipotency and normal differentiation of single teratocarcinoma cells cloned by injection into blastocysts. Proc Natl Acad Sci USA 73(2), 549-553. 

A disaccharide unit with an a-galactose moiety, GaI(al-3)GalNAc has been discovered as an O-linked unit in the brains of the rat, rabbit, hen, frog and fish [31], It seems to be concentrated in nervous tissue, because it was not found in five other tissues of the rat. Other sources where the disaccharide has been discovered are the muscle and the adrenal medulla [32] and human teratocarcinoma cells [33]. Its enrichment in brain tissue together with its occurrence in roughly similar amounts in different animal species may suggest a role important for nervous tissue function. However, no information of its possible role or occurrence in specified glycoprotein molecules is yet available. 

The poly-iV-acetyllactosamine glycans were first discovered as constituents of N-glycans in erythrocytes [36] and teratocarcinoma cells [37], in which they represent a major fraction of the cell surface carbohydrates. In contrast, analysis of whole brain tissue does not indicate the presence of major amounts of poly-77-acetyllactosamine chains [15]. That poly-A -acetyllactosamine glycans do exist in brain tissue is suggested by their accumulation in GMl-gangliosidosis [38]. Small amounts are also present in the form of keratan sulfate as 0-mannose linked chains (see above). 

TYR03 gene processed pseudogene TYR03P are cloned from human teratocarcinoma cell, bone marrow and melanocyte cDNA libraries [537])... 

Stem cell research began in the 1970s with studies on teratocarcinoma cells, which are found in testicular cancers. These cells are bizarre blends of differentiated and undifferentiated cells. They were referred to as embryonal carcinoma (EC) cells. They were found to be pluripotent, which led to the idea of using them for therapy. However, such research was suspended because the cells had come from tumors, which made their use dangerous, and because they were aneuploid, which means they had the wrong number of chromosomes. 

Andrews P W, Damjanov I, Simon D, et al. (1984). Pluripotent embryonal carcinoma clones derived from the human teratocarcinoma cell line Tera-2. Differentiation in vivo and in vitro. Lab. Invest. 50 147-162. 

Many oleane and ursane triterpenoids are reported to have interesting biological, pharmacological, and medicinal activities similar to those of retinoids and steroids. These include antiinflammatory activity, suppression of tumor promotion, suppression of immunoglobulin synthesis, protection of the liver against toxic injury, induction of collagen synthesis, and induction of differentiation in leukemia or teratocarcinoma cells [197]. 

Several other assays of retinoid activity exist [83] and include evaluations of the ability of a retinoid to inhibit the exponential growth of S91 murine melanoma cells or to induce terminal differentiation of the fully neoplastic F9 teratocarcinoma cells. These and other assays of retinoid activity have been useful but the TOC, ODC, and HL-60 assays have been used most comprehensively in establishing retinoid structure-activity relationships, which are discussed below. 

Recent studies by Prehm (1983a, b) have done much to solve the problems of hyaluronate chain assembly and in so doing, he has shown a new mechanism of saccharide synthesis, which may stimulate a re-evaluation of the supposed mechanisms by which some other polysaccharides are constructed. Using a line of teratocarcinoma cells (F9) in which the synthesis of hyaluronate could be induced with retinol, Prehm (1983a) examined the prop-... 

Prehm, P. (1983a) Synthesis of Hyaluronate in differentiated Teratocarcinoma Cells. 

Ohashi Y, Ueda K, Hayaishi O et al. Induction of murine teratocarcinoma cell differentiation by suppression of poly(ADP-ribose) synthesis. Proc Natl Acad Sci USA 1984 81(22) 7132-7136. 

---