An efficient optimized method for isolation of villous trophoblast cells from human early pregnancy placenta suitable for functional and molecular studies.
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.
Stenqvist
Ann-Christin
AC
Department of Clinical Immunology, Umeå University, Umeå, Sweden.
Chen
Ting
T
Hedlund
Malin
M
Dimova
Tanya
T
Nagaeva
Olga
O
Kjellberg
Lennart
L
Innala
Eva
E
Mincheva-Nilsson
Lucia
L
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*
D012333:RNA, Messenger