title: An efficient optimized method for isolation of villous trophoblast cells from human early pregnancy placenta suitable for functional and molecular studies.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Clinical Immunology, Umeå University, Umeå, Sweden.
authors.forename: Ann-Christin
authors.initials: AC
authors.lastname: Stenqvist
authors.forename: Ting
authors.initials: T
authors.lastname: Chen
authors.forename: Malin
authors.initials: M
authors.lastname: Hedlund
authors.forename: Tanya
authors.initials: T
authors.lastname: Dimova
authors.forename: Olga
authors.initials: O
authors.lastname: Nagaeva
authors.forename: Lennart
authors.initials: L
authors.lastname: Kjellberg
authors.forename: Eva
authors.initials: E
authors.lastname: Innala
authors.forename: Lucia
authors.initials: L
authors.lastname: Mincheva-Nilsson
mesh_terms: D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014327:Trophoblasts / Q000166:cytology*
chemical_list: D012333:RNA, Messenger