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

• Published in: Reproductive & developmental biology Vol. 35; no. 3
• Main Authors: Kim, D.H., National Institute of Animal Science, RDA, Suwon, Republic of Korea, Lee, Y.S., University of Tokyo, Tokyo, Japan, Oh, K.B., National Institute of Animal Science, RDA, Suwon, Republic of Korea, Hwang, S.S., National Institute of Animal Science, RDA, Suwon, Republic of Korea, Im, G.S., National Institute of Animal Science, RDA, Suwon, Republic of Korea, Park, J.K., National Institute of Animal Science, RDA, Suwon, Republic of Korea
• Format: Journal Article
• Language: Korean
• Published: 01.09.2011
• Subjects: MICE; Micromanipulation medium; Mouse cloning; Oocyte donor age; RATON; SOURIS
• ISSN: 1738-2432

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 SrCl₂ in Ca-free CZB medium in the presence of 5 ㎍/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 less than 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 less than 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 less than 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.