Overcoming the 2-cell block by modifying standard components in a mouse embryo culture medium.
The 2-cell block may be caused by inappropriate concentrations of commonly used constituents of embryo culture media. Almost all zygotes obtained by fertilizing CF1 ova with hybrid B6D2F1/CrlBR sperm did not develop beyond the 2-cell stage when cultured in Whittingham's medium M16. This 2-cell block was overcome by lowering the concentrations of NaCl, KCl, KH2PO4, glucose, and pyruvate, either individually or in combination. The effects of changing the concentration of either NaCl or KCl depend on the concentration of NaHCO3 in the medium. Although a high percentage of embryos grew to the 4-cell stage in several media with lowered concentrations of certain components, the media are not optimal for complete preimplantation embryo development since the yield of blastocysts is low.
Lawitts
J A
JA
Laboratory of Human Reproduction and Reproductive Biology, Harvard Medical School, Boston, Massachusetts 02115.
Biggers
J D
JD
D000818:Animals; D001639:Bicarbonates; D001755:Blastocyst; D002970:Cleavage Stage, Ovum / Q000166:cytology / Q000187:drug effects*; D003470:Culture Media / Q000032:analysis / Q000494:pharmacology*; D005314:Embryonic and Fetal Development / Q000187:drug effects*; D005260:Female; D005947:Glucose; D008297:Male; D051379:Mice / Q000196:embryology*; D009924:Organ Culture Techniques; D009994:Osmolar Concentration; D011773:Pyruvates; D019289:Pyruvic Acid; D012492:Salts
D001639:Bicarbonates; D003470:Culture Media; D011773:Pyruvates; D012492:Salts; D019289:Pyruvic Acid; D005947:Glucose