Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Comparison of Developmental Competency of Porcine Embryos Cloned with Mesenchymal Stem Cells and Somatic Cells

  • Jin Hai-Feng (Institute of Animal Medicine, College of Veterinary Medicine) ;
  • Kumar B. Mohana (Institute of Animal Medicine, College of Veterinary Medicine) ;
  • Cho Sung-Keun (Division of Applied Life Science, Gyeongsang National University) ;
  • Ock Sun-A (Division of Applied Life Science, Gyeongsang National University) ;
  • Jeon Byeong-Gyun (Division of Applied Life Science, Gyeongsang National University) ;
  • Balasubramanian S. (Department of Clinics, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University) ;
  • Choe Sang-Yong (Institute of Animal Medicine, College of Veterinary Medicine) ;
  • Rho Gyu-Jin (Institute of Animal Medicine, College of Veterinary Medicine)
  • Published : 2006.06.01
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Abstract

The present study compared the developmental potential of cloned porcine embryos with mesenchymal stem cells (MSCs), fetal fibroblasts (FFs) and cumulus cells (CCs) by assessing the cleavage and blastocyst rate, total cell number, inner cell mass (ICM) ratio and apoptosis. MSCs were isolated by ficoll gradients from femur of -6 month old female pig, and maintained for primary cultures. FFs from a female fetus at ${\sim}30$ day of gestation were established, and CCs were obtained from cumulus oocyte complexes (COCs) aspirated from $3{\sim}6$ mm follicles in diameter. Donor cells at $3{\sim}4$ passage were employed for nuclear transfer (NT). COCs were matured and fertilized in vitro(IVF) as control. Cleavage rate was significantly (P<0.05) higher in IVF than in NT embryos with MSCs, FFs and CCs ($82.7{\pm}8.9%\;vs\;70.6{\pm}5.4,\;68.7{\pm}5.1\;and\;63.4{\pm}5.6%$, respectively). However, blastocyst rates in IVF and NT embryos derived from MSCs ($24.5{\pm}2.8\;and\;20.4{\pm}8.3%$) did not differ, but were significantly (P<0.05) higher than NT derived from FFs and CCs ($10.6{\pm}2.7\;and\;9.8{\pm}2.1%$). Total cell number and the ratio of ICM to total cells among blastocysts cloned from MSCs ($35.4{\pm}5.2\;and\;0.40{\pm}0.09%$, respectively) were significantly (P<0.05) higher than those from FFs and CCs ($24.9{\pm}6.2%\;vs\;0.19{\pm}0.16,\;23.6{\pm}5.5\;and\;0.17{\pm}0.16%$, respectively). Proportions of TUNEL positive cells in NT embryos from FFs and CCs ($6.9{\pm}1.5\;and\;7.4{\pm}1.7%$, respectively) were significantly (P<0.05) higher than in MSCs ($4.8{\pm}1.4%$) and IVF ($2.3{\pm}0.9%$). The results demonstrate that MSCs have a greater potential as donor cells than FFs and CCs in achieving enhanced production of cloned porcine embryos.

Keywords

References

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