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The Effect of Oocyte Donor Age and Micromainpulation Medium on the Development of Mouse Cloned Embryos  

Kim, Dong-Hoon (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Lee, Youn-Su (Laboratory of Stem Cell Therapy, Center for Experimental Medicine, Institute of Medical Science, University of Tokyo)
Oh, Keon-Bong (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Hwang, Seong-Soo (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Im, Gi-Sun (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Park, Jin-Ki (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Publication Information
Reproductive and Developmental Biology / v.35, no.3, 2011 , pp. 313-317 More about this Journal
Abstract
This study was conducted to examine the effect of oocyte donor age and micromanipulation medium on the development of mouse cloned embryos receiving cumulus cells. Mouse oocytes were obtained from 6 to 11 week-old mice BDF1 female mice(experiment 1) and cumulus cells were used as donor cells. Micromanipulation procedures for nuclear transfer(NT) were performed in FHM, M2 or Hepes-buffered TCM199(TCM199) medium(experiment 2). After nuclear transfer, the reconstructed oocytes were activated by 10 mM $SrCl_2$ in Ca-free CZB medium in the presence of 5 II ${\mu}$g/ml cytochalasin B for 5 h and cultured in KSOM medium for 4 days. In experiment 1, the survival rate of oocytes after injection of cumulus cells were significantly(p<0.05) lower in oocytes from 6~7 week-old mice(53.3%) than in oocytes from 8~9(80.9%) and 10~11 week-old mice(77.1%). In experiment 2, the survival rate of oocytes after cell injection were significantly(p<0.05) higher in FHM and M2 medium(71.7% and 76.9%) than in TCM199 medium(51.2%). The activation rates of cloned embryos were not different among the micromanipulation media. However, the embryos developed to blastocyst stage were significantly(p<0.05) higher in FHM medium(13.9%) than in M2 and TCM199 medium(0.0% and 0.0%). In conclusion, the present study suggest that oocytes from above 8 week-old mice are superior to oocytes from 6~7 week-old mice as a source of recipient cytoplasm and FHM is superior to M2 and TCM199 as a micromanipulation medium for mouse somatic cell cloning.
Keywords
Mouse cloning; Micromanipulation medium; Oocyte donor age;
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