Storage system comprising an emptied zona pellucida and spermatozoa placed therein and a method of cryopreservation

摘要

A procedure for cryopreservation and efficient post-thaw recovery of a single or few cells, preferably human spermatozoa, their precursor cells or blastomeres, which is achieved by injecting cells into cell- free mammalian zonae pellucidae. The cells may also be of any other type where there are provided a relatively small number of valuable cells which must be retrieved. The zona pellucida may be cryopreserved with the cells contained therein. The method involves a combination of physical micromanipulation procedures and cryoprotectant mediated cryopropreservation. The zonae may be tracked by positioning them into straws between two small air bubbles prior to preservation. There is a high recovery of zonae and sperm within the zonae, and a high fertilization rate of the sperm using ICSI after recovery.