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Human Reproduction, Vol. 15, No. 1, 145-156, January 2000
© 2000 European Society of Human Reproduction and Embryology

Formulation of a protein-free medium for human assisted reproduction*

J. Ali1,2,3, M.A.M. Shahata1 and S.D. Al-Natsha1

1 IVF Unit, Women's Hospital, Hamad Medical Corporation, P.O. Box 3050, Doha, Qatar and 2 JA Global Pty Ltd, P.O. Box 151, Curtin, ACT 2605, Australia

The optimal concentrations of individual amino acids, antioxidants, vitamins, osmolytes and energy sources were determined using a 1-cell Swiss outbred (SO) and or F1 [(CBAxC57BL/6J)xSO] mouse assay in Earle's balanced salt solution containing bovine serum albumin. Based on the findings of these experiments, a number of media were formulated. Of these, the medium showing optimal embryo development and a significantly higher blastocyst hatching rate was investigated further. A protein-free medium (ART-7) was formulated and assessed using 1-, 2- and 4-cell SO mouse embryos. The generation of viable human embryos in the ART-7 series of media in micro- and ultra micro-droplet culture under oil with and without cumulus co-culture following intracytoplasmic sperm injection (ICSI) was investigated. The quality of sibling day 2 human embryos generated in the ART-7 media series was statistically comparable to or better than control embryos. The ART-7 medium was not toxic to human spermatozoa. Fertilization by conventional IVF and subsequent embryo development was not affected. A clinical trial of ICSI-derived embryos generated in the protein-free medium, with and without cumulus co-culture, has resulted in clinical pregnancies (10 of 20 transfers) of which two have proceeded to term, and the remaining patients are in various stages of pregnancy.

Key words: co-culture/embryos/live birth/micro-droplet/protein-free medium

3 To whom correspondence should be addressed at: IVF Unit, Women's Hospital, Hamad Medical Corporation, P.O. Box 3050, Doha, Qatar

* This report was presented in part at the XVI Annual Scientific Meeting of the Fertility Society of Australia, 2–4 December 1997, Adelaide, Australia and the 5th Middle East Fertility Society Annual Meeting, November 18–20, 1998, Amman, Jordan.


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