Abstract
The biology of trophoblast cell development can be investigated using in vitro model systems. The Rcho-1 trophoblast stem cell line was derived from a rat choriocarcinoma and is an effective tool for elucidating regulatory mechanisms controlling trophoblast cell differentiation. In this chapter, we describe methods used in the maintenance and manipulation of the Rcho-1 trophoblast cell line.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Gardner, R. L. and Beddington, R. S. P. (1988) Multi-lineageā stem cellsā in the mammalian embryo. J. Cell Sci., Suppl. 10, 11ā27.
Tanaka, S., Kunath, T., Hadjantonakis, A. K., Nagy, A., and Rossant, J. (1998) Promotion of trophoblast stem cell proliferation by FGF4. Science 282, 2072ā2075.
Rossant, J. (2001) Stem cells from the mammalian blastocyst. Stem Cells 19, 477ā482.
Soares, M. J.,Chapman, B. M., Rasmussen, C. A., Dai, G., Kamei, T., and Orwig, K. E. (1996). Differentiation of trophoblast endocrine cells. Placenta 17, 277ā289.
Ain, R., Canham, L. N., and Soares, M. J. (2003) Gestation stage-dependent intrauterine trophoblast cell invasion in the rat and mouse: novel endocrine phenotype and regulation. Dev. Biol. 260, 176ā190.
Rossant, J. and Cross, J. C. (2001) Placental development: lessons from mouse mutants. Nat. Rev. Genet. 2, 538ā548.
Teshima, S., Shimosato, Y., Koide, T., Kuroki, M., Kikuchi, Y., and Aizawa, M. (1983) Transplantable choriocarcinoma of rats induced by fetectomy and its biological activities. Gann. 74, 205ā212.
Verstuyf, A., Sobis, H., Goebels, J., Fonteyn, E., Cassiman, J. J., and Vandeputte, M. (1990) Establishment and characterization of a continuous in vitro line from a rat choriocarcinoma. Int. J. Cancer 45, 752ā756.
Faria, T. N. and Soares, M. J. (1991) Trophoblast cell differentiation: establishment, characterization, and modulation of a rat trophoblast cell line expressing members of the placental prolactin family. Endocrinology 129, 2895ā2906.
Verstuyf, A., Sobis, H., and Vandeputte, M. (1989) Morphological and immunological characteristics of a rat choriocarcinoma. Int. J. Cancer 44, 879ā884.
Faria, T.N., Deb, S., Kwok, S. C. M., Vandeputte, M., Talamantes, F., and Soares, M. J. (1990) Transplantable rat choriocarcinoma cells express placental lactogen: identification of placental lactogen-I immunoreactive protein and messenger ribonucleic acid. Endocrinology 127, 3131ā3137.
Verstuyf, A., Goebels, J., Sobis, H., and Vandeputte, M. (1993) Influence of different growth factors on a rat choriocarcinoma cell line. Tumour Biol. 14, 46ā54.
Hamlin, G. P. and Soares, M. J. (1995) Regulation of DNA synthesis in proliferating and differentiating trophoblast cells: involvement of transferrin, transforming growth factor-Ī², and tyrosine kinases. Endocrinology 136, 322ā331.
MacAuley, A., Cross, J. C., and Werb, Z. (1998) Reprogramming the cell cycle for endoreduplication in rodent trophoblast cells. Mol. Biol. Cell. 9, 795ā807.
Hattori, N., Davies, T. C., Anson-Cartwright, L., and Cross, J. C. (2000) Periodic expression of the cyclin-dependent kinase inhibitor p57(Kip2) in trophoblast giant cells defines a G2-like gap phase of the endocycle. Mol. Biol. Cell. 11, 1037ā1045.
Balzarini, J., Verstuyf, A., Hatse, S., et al. (1995) The human immunodeficiency virus (HIV) inhibitor 9-(2-phosphonylmethoxyethyl)adenine (PMEA) is a strong inducer of differentiation of several tumor cell lines. Int. J. Cancer 61, 130ā137.
Cross, J. C., Flannery, M. L., Blanar, M. A., et al. (1995) Hxt encodes a basic helix-loop-helix transcription factor that regulates trophoblast cell development. Development 121, 2513ā2523.
Yamaguchi, M., Kawai, M., Kishi, K., and Miyake, A. (1995) Regulation of rat placental lactogen (rPL)-II secretion: cAMP inhibits rPL-secretion in vitro. Eur. J. Endocrinol. 133, 342ā346.
Grummer, R., Hellmann, P., Traub, O., Soares, M. J., and Winterhager, E. (1996) Regulation of connexin 31 gene expression upon retinoic acid treatment in rat choriocarcinoma cells. Exp. Cell Res. 227, 23ā32.
Kamei, T., Hamlin, G. P., Chapman, B. M., Burkhardt, A. L., Bolen, J. B., and Soares, M. J. (1997) Signaling pathways controlling trophoblast cell differentiation: Src family protein tyrosine kinases in the rat. Biol. Reprod. 57, 1302ā1311.
Yamamoto, T., Chapman, B. M., and Soares, M. J. (1997) Protein kinase C dependent and independent mechanisms controlling rat trophoblast cell DNA synthesis and differentiation. J. Reprod. Fertil. 111, 15ā20.
Hatse, S., Naesens, L., De Clercq, E., and Balzarini, J. (1998) Potent differentiation-inducing properties of the antiretroviral agent 9-(2-phosphonylmethoxyethyl)adenine (PMEA) in the rat choriocarcinoma (RCHO) tumor cell model. Biochem. Pharmacol. 56, 851ā859
Nakayama, H., Scott, I. C., and Cross, J. C. (1998) The transition to endoreduplication in trophoblast giant cells is regulated by the mSNA zinc finger transcription factor. Dev. Biol. 199, 150ā163.
Kraut, N., Snider, L., Chen, C. M., Tapscott, S. J., and Groudine, M. (1998) Requirement of the mouse I-mfa gene for placental development and skeletal patterning. EMBO J. 17, 6276ā6288.
Hatse, S., Naesens, L., De Clercq, E., and Balzarini, J. (1999) N6-cyclopropyl-PMEDAP: a novel derivative of 9-(2-phenylphonylmethoxyethyl)-2,6-diaminopurine (PMEDAP) with distinct metabolic, an iproliferative, and differentiation-inducing properties. Biochem. Pharmacol. 58, 311ā323.
Peters, T. J., Chapman, B. M., Wolfe, M. W., and Soares, M. J. (2000) Placental lactogen-I gene activation in differentiating trophoblast cells: extrinsic and intrinsic regulation involving mitogen-activated protein kinase signaling pathways. J. Endocrinol. 165, 443ā456.
Scott, I.C., Anson-Cartwright, L., Riley, P., Reda, D., and Cross, J.C. (2000) The Hand1 basic helix-loop-helix transcription factor regulates trophpblast giant cell differentiation via multiple mechanisms. Mol. Cell Biol. 20, 530ā541.
Ma, G. T., Soloveva, V., Tzeng, S. J., et al. (2001) Nodal regulates trophoblast differentiation and placental development. Dev Biol. 236, 124ā135.
Parast, M. M., Aeder, S., and Sutherland, A. E. (2001) Trophoblast giant-cell differentiation involves changes in cytoskeleton and cell motility. Dev. Biol. 230, 43ā60.
Kamei, T., Jones, S. R., Chapman, B. M., McGonigle, K., Dai, G., and Soares, M. J. (2002) Activation and involvement of the phosphatidylinositol 3-kinase/aktsignaling pathway in the endocrine differentiation of trophoblast cells. Mol. Endocrinol. 16, 1469ā1481.
Hinck, L., Thissen, J. P., Pampfer, S., and De Hertogh, R. (2003) Effect of high concentrations of glucose on differentiation of rat trophoblast cells in vitro. Diabetologia 46, 276ā283.
Takahashi, Y., Carpino, N., Cross, J. C., Torres, M., Parganas, E., and Ihle, J. N. ( 2003) SOCS 3: an essential regulator of LIF receptor signaling in trophoblast giant cell differentiation. EMBO J. 22, 372ā384.
Verstuyf, A., Fonteyn, E., Sobis, H., and Vandeputte, M. (1992) A rat cytotrophoblast antigen defined by a monoclonal antibody. Am. J. Reprod. Immunol. 28, 6ā11.
Hamlin, G. P., Lu, X.-J., Roby, K. F., and Soares, M. J. (1994) Recapitulation of the pathway for trophoblast giant cell differentiation in vitro: stage-specific expression of members of the prolactin gene family. Endocrinology 134, 2390ā2396.
Yamamoto, T., Roby, K. F., Kwok, S. C. M., and Soares, M. J. (1994) Transcriptional activation of cytochrome P450 side chain cleavage enzyme expression during trophoblast cell differentiation. J. Biol. Chem. 269, 6517ā6523.
Yamamoto, T., Chapman, B. M., Clemens, J. W., Richards, J. S., and Soares, M. J. (1995) Analysis of cytochrome P450 side-chain cleavage gene promoter activation during trophoblast cell differentiation. Mol. Cell. Endocrinol. 113, 183ā194.
Yamamoto, T., Chapman, B. M., Johnson, D. C., Givens, C. R., Mellon, S. H., and Soares, M. J. (1996) Cytochrome P450 17Ī±-hydroxylase gene expression in differentiating rat trophoblast cells. J. Endocrinol. 150, 161ā168.
Dai, G., Imagawa, W., Liu, B., et al. (1996) Rcho-1 trophoblast cell placental lactogens: complementary DNAs, heterologous expression, and biological activities. Endocrinology 137, 5020ā5027.
Shirai, T., Itonori, S., Soares, M. J., Shiota, K., and Ogawa, T. (1996) Changes in ganglioside composition of the rat choriocarcinoma cell line, Rcho-1. Glycoconjugate J. 13, 415ā421.
Chun, J. Y., Han, Y. J., and Ahn, K. Y. (1999) Psx homeobox gene is X-linked and specifically expressed in trophoblast cells of mouse placenta. Dev. Dyn. 216, 257ā266.
Hwang, I. T., Lee, Y. H., Moon, B. C., Ahn, K. Y., Lee, S. W., and Chun, J. Y. (2000) Identification and characterization of a new member of the placental prolactin-like protein-C (PLP-C) subfamily, PLP-CĪ². Endocrinology 141, 3343ā3352.
Sahgal, N., Knipp, G. T., Liu, B., Chapman, B. M., Dai, G., and Soares, M. J. (2000) Identification of two new nonclassical members of the rat prolactin family. J. Mol. Endocrinol. 24, 95ā108.
Oda, M., Sun, W., Hattori, N., Tanaka, S., and Shiota, K. (2001) PAL31 expression in rat trophoblast giant cells. Biochem. Biophys. Res. Commun. 287, 721ā726.
Dai, G., Lu, L., Tang, S., Peal, M. J., and Soares, M. J. (2002) The prolactin family miniarray: a tool for evaluating uteroplacental/trophoblast endocrine cell phenotypes. Reproduction 124, 755ā765.
Mehta, D. V., Kim, Y. S., Dixon, D., and Jetten, A. M. (2002) Characterization of the expression of the retinoid-related, testis-associated receptor (RTR) in trophoblasts. Placenta 23, 281ā287.
Morris Buus, R. and Boockfor, F. R. (2004) Transferrin expression by placental trophoblastic cells. Placenta 25, 45ā52.
Novak, D., Quiggle, F., and Matthews, J. (2004) Androgen secretion by Rcho-1 cells is independent of extracellular glutamate concentration. Placenta 25, 548ā552.
Shida, M. M., Ng, Y.-K., Soares, M. J., and Linzer, D. I. H. (1993) Trophoblast-specific transcription from the mouse placental lactogen I gene promoter. Mol Endocrinol. 12, 181ā188.
Vuille, J. C., Cattini, P. A., Bock, M. E., et al. (1993) Rat prolactin-like protein A partial gene and promoter structure: promoter activity in placental and pituitary cells. Mol. Cell. Endocrinol. 96, 91ā98.
Ng, Y. K., George, K. M., Engel, J. D., and Linzer, D. I. H. (1994) GATA factor activity is required for the trophoblast-specific transcriptional regulation of the mouse placental lactogen I gene. Development 120, 3257ā3266.
Dai, G., Liu, B., Szpirer, C., Levan, G., Kwok, S. C. M., and Soares, M. J. (1996) Prolactin-like protein-C variant: complementary DNA, unique six exon gene structure, and trophoblast cell-specific expression. Endocrinology 137, 5009ā5019.
Scatena, C. D. and Adler, S. (1996) Trans-acting factors dictate the species-specific placental expression of corticotropin-releasing factor genes in choriocarcinoma cell lines. Endocrinology 137, 3000ā3008.
Orwig, K. E., Dai, G., Rasmussen, C. A., and Soares, M. J. (1997) Decidual/trophoblast prolactin-related protein: characterization of gene structure and cell-specific expression. Endocrinology 138, 2491ā2500.
Lin, J. and Linzer, D. I. H. (1998) Identification of trophoblast-specific regulatory elements in the mouse placental lactogen II gene. Mol. Endocrinol. 12, 418ā427.
Matsumoto, K., Yamamoto, T., Kurachi, H., et al. (1998) Human chorionic gonadotropin-Ī± gene is transcriptionally activated by epidermal growth factor through cAMP response element in trophoblast cells. J. Biol. Chem. 273, 7800ā7806.
Shah, P., Sun, Y., Szpirer, C., and Duckworth, M. L. (1998) Rat placental lactogen II gene: characterization of gene structure and placental expression. Endocrinology 139, 967ā973.
Sun, Y., Robertson, M. C., and Duckworth, M. L. (1998) The effects of epidermal growth factor/transforming growth factor-Ī± on the regulation of placental lactogen I and II mRNAs in a rat choriocarcinoma cell line. Endocr. J. 45, 297ā306.
Dai, G., Wolfe, M. W., and Soares, M. J. (1999) Distinct regulatory regions from the prolactin-like protein-C variant promoter direct trophoblast giant cell versus spongiotrophoblast cell-specific expression. Endocrinology 140, 4691ā4698.
Liang, R., Limesand, S. W., and Anthony, R. V. (1999) Structure and transcriptional regulation of the ovine placental lactogen gene. Eur. J. Biochem. 265, 883ā895.
Orwig, K. E. and Soares, M. J. (1999) Transcriptional activation of the decidual/trophoblast prolactin-related protein gene. Endocrinology 140, 4032ā4039.
Sun, Y. and Duckworth, M. L. (1999) Identification of a placental-specific enhancer in the rat placental lactogen II gene that contains binding sites for members of the Ets and AP-1 (activator protein 1) families of transcription factors. Mol. Endocrinol. 13, 385ā399.
Okamoto, Y., Sakata, M., Yamamoto, T., et al. (2001) Involvement of nuclear transcription factor Sp1 in regulating glucose transporter-1 gene expression during rat trophoblast differentiation. Biochem. Biophys. Res.Commun. 288, 940ā948.
Yamamoto, T., Matsumoto, K., Kurachi, H., et al. (2001) Progesterone inhibits transcriptional activation of human chorionic gonadotropin-alpha gene through protein kinase A pathway in trophoblast cells. Mol. Cell. Endocrinol. 182, 215ā224.
Szafranska, B., Miura, R., Ghosh, D., et al. (2001) Gene for porcine pregnancy-associated glycoprotein 2 (poPAG2): its structural organization and analysis of its promoter. Mol. Reprod. Dev. 60, 137ā146.
Gabriel, H. D., Strobl, B., Hellmann, P., Buettner, R., and Winterhager, E. (2001) Organization and regulation of the rat Cx31 gene. Implication for a crucial role of the intron region. Eur. J. Biochem. 268, 1749ā1759.
Firulli, B. A., Howard, M. J., McDaid, J. R., et al. (2003) PKA, PKC, and the protein phosphatase 2A influence HAND factor function: a mechanism for tissue-specific transcriptional regulation. Mol. Cell 12, 1225ā1237.
Limesand, S. W., Jeckel, K. M., and Anthony, R. V. (2004) Pur alpha, a single-stranded deoxyribonucleic acid binding protein, augments placental lactogen gene transcription. Mol. Endocrinol. 18, 447ā457.
Hershberger M. E. and Tuan, R. S. (1998) Placental 57-kDa Ca(2+)-binding protein: regulation of expression and function in trophoblast calcium transport. Dev. Biol. 199, 80ā92.
Hershberger, M. E. and Tuan, R. S. (1999) Functional analysis of placental 57-kDa Ca(2+)-binding protein: overexpression and downregulation in a trophoblastic cell line. Dev. Biol. 215, 107ā117.
Knipp, G. T., Liu, B., Audus, K. L., Fujii, H., Ono, T., and Soares, M. J. (2000) Fatty acid transport regulatory proteins in the developing rat placenta and in trophoblast cell culture models. Placenta 21, 367ā375.
Evans, T. J., James-Kracke, M. R., Kleiboeker, S. B., and Casteel, S. W. (2003) Lead enters Rcho-1 trophoblastic cells by calcium transport mechanisms and complexes with cytosolic calcium-binding proteins. Toxicol. Appl. Pharmacol. 186, 77ā89.
Novak, D. and Matthews, J. (2003) Glutamate transport by Rcho-1 cells derived from rat placenta. Pediatr. Res. 53, 1025ā1029.
Kimura, H., Takeda, T., Tanaka, S., Ogawa, T., and Shiota, K. (1998) Expression of rat DNA (cytosine-5) methyltransferase (DNA MTase) in rodent trophoblast giant cells: molecular cloning and characterization of rat DNA MTase. Biochem. Biophys. Res. Commun. 253, 495ā501.
Ohgane, J., Hattori, N., Oda, M., Tanaka, S., and Shiota, K. (2002) Differentiation of trophoblast lineage is associated with DNA methylation and demethylation. Biochem. Biophys. Res. Commun. 290, 701ā706.
Peters, T. J., Albieri, A., Bevilaqua, E., et al. (1999) Differentiation-dependant expression of gelatinase B/ matrix metalloproteinase-9 in trophoblast cells. Cell Tiss. Res. 295, 287ā296.
Hatse, S., Naesens, L., Degreve, B., et al. (1998) Potent antitumor activity of the acyclic nucleoside phosphonate 9-(2-phosphonylmethoxyethyl)adenine in choriocarcinoma-bearing rats. Int. J. Cancer 76, 595ā600.
Naesens, L., Hatse, S., Segers, C., et al. (1999) 9-(2-phosphonylmethoxyethyl)-N6-cyclopropyl-2,6-diaminopurine: a novel prodrug of 9-(2-phosphonylmethoxyethyl)guanine with improved antitumor efficacy and selectivity in choriocarcinoma-bearing rats. Oncol. Res. 11, 195ā203.
Pierce, G. B. (1985) Carcinoma is to embryology as mutation is to genetics. Am. Zool. 25, 707ā712.
Rudnicki, M. A. and McBurney, M. W. (1987) Cell culture methods and induction of differentiation of embryonal carcinoma cell lines, in Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, IRL, Oxford, UK: pp. 19ā29.
Duckworth, M. L., Schroedter, I. C., and Friesen, H. G. (1990) Cellular localization of rat placental lactogen-II and rat prolactin-like proteins A and B by in situ hybridization. Placenta 11, 143ā155.
Cohick, C. B., Xu, L., and Soares, M. J. (1997) Prolactin-like protein-B: heterologous expression and characterization of placental and decidual species. J. Endocrinol. 152, 291ā302.
Iwatsuki, K. Shinozaki, M., Sun, W., Yagi, S., Tanaka, S., and Shiota, K. (2000) A novel secretory protein produced by rat spongiotrophoblast. Biol. Reprod. 62, 1352ā1359.
Toft, D. J. and Linzer, D. I. (2000) Identification of three prolactin-related hormones as markers of invasive trophoblasts in the rat. Biol. Reprod. 63, 519ā525.
Wiemers, D. O., Ain, R., Ohboshi, S., and Soares, M. J. (2003) Migratory trophoblast cells express a newly identified member of the prolactin gene family. J. Endocrinol. 179, 335ā346.
Tomogane, H., Mistry, A. M., and Voogt, J. L. (1991) Late pregnancy and rat choriocarcinoma cells inhibit nocturnal prolactin surges and serotonin-induced prolactin release. Endocrinology 130, 23ā28.
Arbogast, L. A., Soares, M. J., Tomogane, H., and Voogt, J. L. (1992) A trophoblast-specific factor(s) suppress circulating prolactin levels and increases tyrosine hydroxylase activity in tuberoinfundibular dopaminergic neurons. Endocrinology 131, 105ā113.
Mathiasen, J. R., Tomogane, H., and Voogt, J. L. (1992) Serotonin-induced decrease in hypothalamic tyrosine hydroxylase activity and corresponding increase in prolactin release are abolished at midpregnancy and by transplants of rat choriocarcinoma cells. Endocrinology 131, 2527ā2532.
Arbogast, L. A., Soares, M. J., Robertson, M. C., and Voogt, J. L. (1993) A trophoblast-specific factor increases tyrosine hydroxylase activity in fetal hypothalamic cell cultures. Endocrinology 133, 111ā120.
Tomogane, H., Arbogast, L. A., Soares, M. J., Robertson, M. C., and Voogt, J. L. (1993) A factor(s) from a rat trophoblast cell line inhibits prolactin secretion in vitro and in vivo. Biol. Reprod. 48, 325ā332.
Sobis, H., Waer, M., and Vandeputte, M. (1996) Normal and malignant trophoblasts do not recruit granulated metrial gland cells. Tumour Biol. 17, 13ā19.
Flietstra, R. J. and Voogt, J. L. (1997) Lactogenic hormones of the placenta and pituitary inhibit suckling-induced prolactin (PRL) release but not the ante-partum PRL surge. Proc. Soc. Exp. Biol. Med. 214, 258ā264.
Lee, Y. and Voogt, J. L. (1999) Feedback effects of placental lactogens on prolactin levels and Fos-related antigen immunoreactivity of tuberoinfundibular dopaminergic neurons in the arcuate nucleus during pregnancy in the rat. Endocrinology 140, 2159ā2166.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
Ā© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Sahgal, N., Canham, L.N., Canham, B., Soares, M.J. (2006). Rcho-1 Trophoblast Stem Cells. In: Soares, M.J., Hunt, J.S. (eds) Placenta and Trophoblast. Methods in Molecular Medicineā¢, vol 121. Humana Press. https://doi.org/10.1385/1-59259-983-4:157
Download citation
DOI: https://doi.org/10.1385/1-59259-983-4:157
Publisher Name: Humana Press
Print ISBN: 978-1-58829-404-3
Online ISBN: 978-1-59259-983-7
eBook Packages: Springer Protocols