title: [Isolation, culture and identification of sheep multinucleated chorionic trophoblast cells].
abstract: OBJECTIVE
  To develop a set of methods for isolating and culturing sheep multinucleated chorionic trophoblast cells and identify them.
  
  
  METHODS
  Trophoblast cells were isolated and cultured by two-step digestion of trypsin and collagenase, and observed under an inverted phase-contrast microscope. They were further identified through conventional HE staining, immunohistochemical staining and transmission electron microscopy.
  
  
  RESULTS
  Under an inverted phase-contrast microscope, the obtained trophoblast cells exhibited epithelioid and sheet-like spreading growth with binuclear or multinuclear. The multinucleated trophoblast cytoplasm from the sheep placental cotyledons and the slides of the trophoblast cells were all stained brown and exhibited positive reactivity by immunohistochemical staining with cytokeratin antibodies. Also abundance of microvilli on surface of cells, together with intracytoplasmic vacuoles, microfilament and lipid droplets were observed under a transmission electron microscope.
  
  
  CONCLUSIONS
  Two-step digestion of trypsin and collagenase has been established for the isolation and cultivation of sheep multinucleated trophoblast cells, and using it, we obtained sheep multinucleated chorionic trophoblast cells with a high purity and biological activity.
authors.affiliation: College of Veterinary Medicine, Inner Mongolia Agricultural University, Ministry of Agriculture, Huhhot 010018, China.
authors.forename: Zhuanjia
authors.initials: Z
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Inner Mongolia Agricultural University, Ministry of Agriculture, Huhhot 010018, China.
authors.forename: Yufei
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, Inner Mongolia Agricultural University; Key Laboratory of Animal Clinical Diagnosis Technology, Ministry of Agriculture, Huhhot 010018, China.
authors.forename: Shuying
authors.initials: S
authors.lastname: Liu
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002467:Cell Nucleus / Q000378:metabolism*; D002469:Cell Separation / Q000379:methods*; D002823:Chorion / Q000166:cytology*; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D053552:Keratin-7 / Q000378:metabolism; D011247:Pregnancy; D012756:Sheep; D014327:Trophoblasts / Q000166:cytology*
chemical_list: D053552:Keratin-7