Skip to main content

In Vitro Models for Studying Trophoblast Transcellular Transport

  • Protocol
Placenta and Trophoblast

Part of the book series: Methods in Molecular Medicineā„¢ ((MIMM,volume 122))

Abstract

In vitro models have proven to be effective in studying the placental transporters that play a role in the exchange of nutrients, waste products, and drugs between the maternal and fetal circulations. Although primary cultures of trophoblast cells can be used to perform uptake, efflux, and metabolism studies, only the rodent HRP-1 and the human BeWo cell lines have been shown to form confluent monolayers when grown on semi-permeable membranes. Protocols for the revival, maintenance, passage, and growth of BeWo cells for transporter expression and transcellular transport studies are provided.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 229.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Stulc, J. (1989) Extracellular transport pathways in the haemochorial placenta. Placenta 10, 113ā€“119.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  2. Sibley, C. P. (1994) Mechanisms of ion transfer by the rat placenta-a model for the human placenta. Placenta 15, 675ā€“691.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  3. Enders, A. C. and Blankenship, T. N. (1999) Comparative placental structure. Adv. Drug Delivery Rev. 38, 3ā€“15.

    ArticleĀ  CASĀ  Google ScholarĀ 

  4. Ringler, G. E. and Strauss, J. F. III (1990) In vitro systems for the study of human placental endocrine function. Endocrine Rev. 11, 105ā€“123.

    ArticleĀ  CASĀ  Google ScholarĀ 

  5. Kliman, H. J., Nestler, J. E., Sermasi, E., Sanger, J. M., and Strauss, J. F. III (1986) Purification, characterization, and in vitro differentiation of cytotrophoblasts from human term placentae. Endocrinology 118, 1567ā€“1582.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  6. Hemmings, D. G., Lowen, B., Sherburne, R., Sawicki, G., and Guilbert, L. J. (2001) Villous trophoblasts cultured on semi-permeable membranes form an effective barrier to the passage of high and low molecular weight particles. Placenta 22, 70ā€“79.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  7. King, A., Thomas, L., and Bischof, P. (2000) Cell culture models of trophoblast II: trophoblast cell lines-a workshop report. Placenta 21, S113ā€“S119.

    ArticleĀ  PubMedĀ  Google ScholarĀ 

  8. Xu, R. H., Chen, X., Li, D. S., Li, R., Addicks, G. C., Glennon, C., Zwaka, T. P., and Thomson, J. A. (2002) BMP4 initiates human embryonic stem cell differentiation to trophoblast. Nat. Biotechnol. 20, 1261ā€“1264.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  9. Tanaka, S., Kunath, T., Hadjantonakis, A. K., Nagy, A., and Rossant, J. (1998) Promotion of trophoblast stem cell proliferation by FGF4. Science 282, 2072ā€“2075.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  10. Liu, F., Soares, M. J., and Audus, K. L. (1997) Permeability properties of monolayers of the human trophoblast cell line BeWo. Am. J. Physiol. 42, C1596ā€“C1604.

    Google ScholarĀ 

  11. Shi, F. L., Soares, M. J., Avery, M., Liu, F., Zhang, X. M., and Audus, K. L. (1997) Permeability and metabolic properties of a trophoblast cell line (HRP-1) derived from normal rat placenta. Exp. Cell Res. 234, 147ā€“155.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  12. 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, 367ā€“375.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  13. Das, U. G., Sadiq, H. F., Soares, M. J., Hay, W. W., and Devaskar, S. U. (1998) Time-dependent physiological regulation of rodent and ovine placental glucose transporter (GLUT-1) protein. Am. J. Physiol. 43, R339ā€“R347.

    Google ScholarĀ 

  14. Rajakumar, R. A., Thamotharan, S., Menon, R. K., and Devaskar, S. U. (1998) Sp1 and Sp3 regulate transcriptional activity of the facilitative glucose transporter isoform-3 gene in mammalian neuroblasts and trophoblasts. J. Biol. Chem. 273, 27,474ā€“27,483.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  15. Novak, D., Quiggle, F., Artime, C., and Beveridge, M. (2001) Regulation of glutamate transport and transport proteins in a placental cell line. Am. J. Physiol. 281, C1014ā€“C1022.

    CASĀ  Google ScholarĀ 

  16. Zhou, F., Tanaka, K., Soares, M. J., and You, G. F. (2003) Characterization of an organic anion transport system in a placental cell line. Am. J. Physiol. 285, E1103ā€“E1109.

    CASĀ  Google ScholarĀ 

  17. Pattillo, R. A. and Gey, G. O. (1968) The establishment of a cell line of human hormone-synthesizing trophoblastic cells in vitro. Cancer Res. 28, 1231ā€“1236.

    CASĀ  PubMedĀ  Google ScholarĀ 

  18. Friedman, S. J. and Skehan, P. (1979) Morphological differentiation of human choriocarcinoma cells induced by methotrexate. Cancer Res. 39, 1960ā€“1967.

    CASĀ  PubMedĀ  Google ScholarĀ 

  19. Wice, B., Menton, D., Geuze, H., and Schwartz, A. L. (1990) Modulators of cyclic AMP metabolism induce syncytiotrophoblast formation in vitro. Exp. Cell Res. 186, 306ā€“316.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  20. Zhao, H. Y. and Hundal, H. S. (2000) Identification and biochemical localization of a Na-K-Cl cotransporter in the human placental cell line BeWo. Biochem. Biophys. Res. Commun. 274, 43ā€“48.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  21. Furesz, T. C., Smith, C. H., and Moe, A. J. (1993) ASC system activity is altered by development of cell polarity in trophoblast from human placenta. Am. J. Physiol. 265, C212ā€“C217.

    CASĀ  PubMedĀ  Google ScholarĀ 

  22. Moe, A. J., Furesz, T. C., and Smith, C. H. (1994) Functional characterization of L-alanine transport in a placental choriocarcinoma cell line (BeWo). Placenta 15, 797ā€“802.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  23. Way, B. A., Furesz, T. C., Schwarz, J. K., Moe, A. J., and Smith, C. H. (1998) Sodium-independent lysine uptake by the BeWo choriocarcinoma cell line. Placenta 19, 323ā€“328.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  24. Shah, S. W., Zhao, H., Low, S. Y., McArdle, H. J., and Hundal, H. S. (1999) Characterization of glucose transport and glucose transporters in the human choriocarcinoma cell line, BeWo. Placenta 20, 651ā€“659.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  25. Vardhana, P. A. and Illsley, N. P. (2002) Transepithelial glucose transport and metabolism in BeWo choriocarcinoma cells. Placenta 23, 653ā€“660.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  26. Schmid, K. E., Davidson, W. S., Myatt, L., and Woollett, L. A. (2003) Transport of cholesterol across a BeWo cell monolayer: implications for net transport of sterol from maternal to fetal circulation. J. Lipid Res. 44, 1909ā€“1918.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  27. Takahashi, T., Utoguchi, N., Takara, A., Yet al. (2001) Carrier-mediated transport of folic acid in BeWo cell monolayers as a model of the human trophoblast. Placenta 22, 863ā€“869.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  28. van der Ende, A., du Maine, A., Schwartz, A. L., and Strous, G. J. (1989) Effect of ATP depletion and temperature on the transferrin-mediated uptake and release of iron by BeWo choriocarcinoma cells. Biochem. J. 259, 685ā€“692.

    PubMedĀ  Google ScholarĀ 

  29. van der Ende, A., du Maine, A., Schwartz, A. L., and Strous, G. J. (1990) Modulation of transferrin-receptor activity and recycling after induced differentiation of BeWo choriocarcinoma cells. Biochem. J. 270, 451ā€“457.

    PubMedĀ  Google ScholarĀ 

  30. Prasad, P. D., Hoffmans, B. J., Moe, A. J., Smith, C. H., Leibach, F. H., and Ganapathy, V. (1996) Functional expression of the plasma membrane serotonin transporter but not the vesicular monoamine transporter in human placental trophoblasts and choriocarcinoma cells. Placenta 17, 201ā€“207.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  31. Eaton, B. M. and Sooranna, S. R. (1998) Regulation of the choline transport system in superfused microcarrier cultures of BeWo cells. Placenta 19, 663ā€“669.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  32. Ellinger, I., Schwab, M., Stefanescu, A., Hunziker, W., and Fuchs, R. (1999) IgG transport across trophoblast-derived BeWo cells: a model system to study IgG transport in the placenta. Eur. J. Immunol. 29, 733ā€“744.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  33. Gao, J.N., Hugger, E.H., Beck-Westermeyer, M.S., and Borchardt, R.T. (2000) Estimating intestinal mucosal permeation of compounds using Caco-2 cell monolayers. Current Protocols in Pharmacology Supplement 8, 7.2.1.ā€“7.2.23.

    Google ScholarĀ 

  34. Silverstein, P. S. Karunaratne, D. N., and Audus, K. L. (2003) Uptake studies for evaluating activity of efflux transporters in a cell line representative of the bloodbrain barrier. Current Protocols in Pharmacology Supplement 23, 7.7.1ā€“7.7.14.

    Google ScholarĀ 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

Ā© 2006 Humana Press Inc.

About this protocol

Cite this protocol

Bode, C.J., Jin, H., Rytting, E., Silverstein, P.S., Young, A.M., Audus, K.L. (2006). In Vitro Models for Studying Trophoblast Transcellular Transport. In: Soares, M.J., Hunt, J.S. (eds) Placenta and Trophoblast. Methods in Molecular Medicineā„¢, vol 122. Humana Press. https://doi.org/10.1385/1-59259-989-3:225

Download citation

  • DOI: https://doi.org/10.1385/1-59259-989-3:225

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-608-5

  • Online ISBN: 978-1-59259-989-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics