The use of an alginic acid matrix to support in vitro development of isolated murine blastomeres

The present study reports the use of a cross-linked calcium alginate matrix to replace the zona pellucida in supporting in vitro development of isolated two-cell murine blastomeres. The zona pellucidae were enzymatically removed with a 0.5% pronase solution and blastomeres separated with a fine-bore glass pipet. The matrix was created by coating the blastomeres with a 3.5% sodium alginic acid solution, which forms a mechanically stable complex upon transfer to media containing high concentrations of a divalent cation. Blastocyst formation was 55/61 (90.2%) for untreated intact embryos and 61/87 (70.1%) and 27/65 (41.5%) for blastomeres cultured with and without the artificial matrix respectively (P less than 0.001).