Success in producing a human embryonic stem cell line depends largely on the quality of the 5-d-old embryo (blastocyst) used. Such blastocysts are grown from frozen-thawed d 1-3 surplus embryos left over from infertility clinics and donated for stem cell research with informed patient consent. Knowledge, therefore, of the culture conditions and embryo scoring systems that are used to generate high-quality blastocysts are crucial. This chapter describes an extended microdroplet culture protocol using two-stage sequential culture media with morphological and polarized optical parameters for embryo scoring for each day of embryonic growth to help produce high-quality blastocysts.
