ORIGINAL ARTICLE: An Efficient Optimized Method for Isolation of Villous Trophoblast Cells from Human Early Pregnancy Placenta Suitable for Functional and Molecular Studies
Article first published online: 28 JUN 2008
© 2008 The Authors. Journal compilation © 2008 Blackwell Munksgaard
American Journal of Reproductive Immunology
Volume 60, Issue 1, pages 33–42, July 2008
How to Cite
Stenqvist, A.-C., Chen, T., Hedlund, M., Dimova, T., Nagaeva, O., Kjellberg, L., Innala, E. and Mincheva-Nilsson, L. (2008), ORIGINAL ARTICLE: An Efficient Optimized Method for Isolation of Villous Trophoblast Cells from Human Early Pregnancy Placenta Suitable for Functional and Molecular Studies. American Journal of Reproductive Immunology, 60: 33–42. doi: 10.1111/j.1600-0897.2008.00588.x
- Issue published online: 28 JUN 2008
- Article first published online: 28 JUN 2008
- Submitted November 26, 2007; accepted January 30, 2008.
- Human placenta;
- isolation method;
- Percoll gradient;
- villous trophoblast
Problem The uniqueness of the human placenta cannot be replaced by animal models. In vitro studies are compulsory to elucidate the biology of human placenta and require isolation and purification of villous trophoblasts, which can be used in molecular and functional studies. Constant improvement in the isolation technique is required to obtain a high yield of pure trophoblast cells with high viability and well preserved morphology.
Method of study Optimized isolation procedure for human villous trophoblasts based on mild enzymatic treatment, Percoll gradient centrifugation and additional purification step involving positive or negative immunoselection on magnetic beads is described.
Results A simple and effective isolation protocol gave a reasonably high yield of villous trophoblast cells with high purity and viability, and excellent morphology as assessed by flow cytometry and electron microscopy.
Conclusion This protocol provides an efficient, optimized method for isolation and enrichment of villous trophoblast cells, suitable for phenotypic, ultrastructural, molecular and functional analyses and for establishment of primary cultures.