Normal in-vivo development of marmoset monkey embryos after trophectoderm biopsy

A procedure was developed to remove trophectoderm cells from day-8 blastocysts of the marmoset monkey. Using micromanipulative techniques, a tear was made in the zona pellucida opposite the inner cell mass which facilitated the controlled herniation of trophectoderm cells as the blastocysts developed in vitro. After 24 h (day-9 blastocysts) and 48 h (day-10 blastocysts) of culture approximately 20% and approximately 50% respectively of the blastocyst had herniated. The herniated trophectoderm was cut off by freehand and the biopsied blastocysts transferred to recipients. Normal offspring were born but pregnancies could be established from day-10 blastocysts only if the recipients were treated with human chorionic gonadotrophin during early pregnancy. One pregnancy was established after the transfer of three frozen biopsied day-10 blastocysts. Biopsies of 30-50 cells from day-10 blastocysts could be readily grown in vitro as trophoblast vesicles to in excess of 1000 cells but biopsies of less than 20 cells from day-9 blastocysts formed a monolayer of binucleated and multinucleated cells with limited cell replication. Assuming human trophectoderm cells have a similar capacity to the marmoset to grow in vitro, the application of this technique to human blastocysts would provide sufficient cells on which the preimplantation diagnosis of a genetic disorder could be made.