Formulation of a protein-free medium for human assisted reproduction.
The optimal concentrations of individual amino acids, antioxidants, vitamins, osmolytes and energy sources were determined using a 1-cell Swiss outbred (SO) and or F(1) [(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.
Ali
J
J
IVF Unit, Women's Hospital, Hamad Medical Corporation, P.O. Box 3050, Doha, Qatar and JA Global Pty Ltd, P.O. Box 151, Curtin, ACT 2605, Australia.
Shahata
M A
MA
Al-Natsha
S D
SD
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D003470:Culture Media; D046508:Culture Techniques; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D011247:Pregnancy; D011506:Proteins; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D020554:Sperm Injections, Intracytoplasmic
D003470:Culture Media; D011506:Proteins; D012709:Serum Albumin; D012710:Serum Albumin, Bovine