Preparation of metaphase spreads from chorionic villus samples

Chorionic villi are composed of an outer layer of trophoblastic cells and an inner mesenchymal cell core, both of fetal origin. Cytogenetic analysis of chorionic villi can be accomplished using material prepared in either of two ways. In the culture method described in this unit, villi are disaggregated by mechanical and enzymatic methods, and the resulting cell suspension is used to establish primary cultures. Mesenchymal cells of the villus core are released by this procedure and the fibroblasts are actively proliferative in tissue culture. Cultures can be used for cytogenetic analysis after 1 week. In the "direct" technique, presented here in an Alternate Protocol, Langhans cells of the cytotrophoblast, actively dividing cells in first-trimester villi, are synchronized and arrested in mitosis after a short incubation period, and metaphase spreads are prepared. Chorionic villi are composed of an outer layer of trophoblastic cells and an inner mesenchymal cell core, both of fetal origin.