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Regular Article |
a Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115
ABSTRACT
Development of outbred CF1 mouse zygotes in vitro was studied in a chemically defined, protein-free medium both with and without amino acids. The addition of amino acids to protein-free potassium simplex optimized medium (KSOM) had little effect on the proportion of embryos that developed at least to the zona-enclosed blastocyst stage. In contrast, amino acids stimulated very significantly, in a dilution-dependent way, the proportion of blastocysts that at least partially or completely hatched. Amino acids also stimulated cell proliferation in both the trophectoderm and inner cell mass (ICM) cells, at rates that favored proliferation of cells in the ICM; had no effect on the incidence of cell death (oncosis or apoptosis); and improved development of the basement membranes, which form on the blastocoelic surface of the trophectoderm and between the primitive endoderm and the primitive ectoderm. Thus, KSOM, supplemented with amino acids but containing no protein supplements, supports development of a newly fertilized ovum to the late blastocyst stage, in which its normal, three-dimensional structure is preserved and in which the ICM has been partitioned into the primitive ectoderm and primitive endoderm.
First decision: 22 October 1999.
1 Supported as part of the National Cooperative Program on Non-Human In Vitro Fertilization and Preimplantation Development and funded by the National Institute of Child Health and Human Development, NIH, through cooperative agreement HD21988.
2 Correspondence: John D. Biggers, Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115. FAX: 617 432 2229; john_biggers{at}hms.harvard.edu
3 Current address: Department of Biology, University of Michigan, Ann Arbor, MI 48109.
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