Abstract
Parthenogenesis is an artificial oocytes activation process without paternal contribution. Blastocyst, derived from parthenogenesis, is one of potential source for pluripotent stem cell propagation. Unfortunately, previous studies reported that parthenogenetic embryo did not achieve exhilarating blastocyst rate. One of the component that predicted inhibit parthenogenetic embryo development is phosphate. Therefore, we try to modify culture medium in order to overcome that problem. The aim of this research was to produce and analyze the characteristics of parthenogenetic blastocyst developed in phosphate-free medium. Mouse oocytes obtained from adult female DDY by superovulation. The activator was strontium chloride 10 mM and diploidization with cytochalasin B 5 μg/ml. Medium for activation and culture medium were modified rat 1 cell embryo medium (MR1ECM) which is phosphate free. The results showed that parthenotes that were cultured in phosphate free medium reached higher blastocyst rate compared to the other groups. The increase of phosphate in culture medium lead to impaired parthenogenetic embryos development. Further experiment was made to analyze the differences between fertilized and parthenogenetic embryo in this medium. The experiment showed that diploid parthenogenetic could achieve high blastocyst rate (30.9±1.3%). The quality of diploid parthenogenetic blastocyst, based on cells number, viability, and ICM ratio, was lower than fertilized blastocyst.
Highlights
Embryonic stem cells (ESCs) comes from early embrional stage known as blastocyst
The results showed that parthenotes that were cultured in phosphate free medium reached higher blastocyst rate compared to the other groups
The aim of this research was to produce and analyze the characteristics of diploid parthenogenetic blastocyst developed in phosphate free medium
Summary
Embryonic stem cells (ESCs) comes from early embrional stage known as blastocyst. At this stage, embryo differentiates into two distinct lineages, trophectoderm and inner cell mass (Kidder 2014). The trophectoderm posseses epithelium characteristics and further develop and form extraembryonal tissue (Marikawa and Alarcon 2009). Inner cell mass potentially capable to give rise to the three primary germ layer that later become all types of body cells. Stem cells derived from inner cell mass have pluripotency properties and can be classified as pluripotent stem cells. Since the first invention of mouse ESCs in 1981 by Evans and Kaufman, method for culturing
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