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ORIGINAL ARTICLE: An Efficient Optimized Method for Isolation of Villous Trophoblast Cells from Human Early Pregnancy Placenta Suitable for Functional and Molecular Studies
Author(s) -
Stenqvist AnnChristin,
Chen Ting,
Hedlund Malin,
Dimova Tanya,
Nagaeva Olga,
Kjellberg Lennart,
Innala Eva,
MinchevaNilsson Lucia
Publication year - 2008
Publication title -
american journal of reproductive immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.071
H-Index - 97
eISSN - 1600-0897
pISSN - 1046-7408
DOI - 10.1111/j.1600-0897.2008.00588.x
Subject(s) - trophoblast , percoll , placenta , biology , flow cytometry , differential centrifugation , microbiology and biotechnology , isolation (microbiology) , centrifugation , andrology , biochemistry , fetus , pregnancy , bioinformatics , medicine , genetics
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.