• Title/Summary/Keyword: 이종 이식

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Effects of Recipient Oocytes and Electric Stimulation Condition on In Vitro Development of Cloned Embryos after Interspecies Nuclear Transfer with Caprine Somatic Cell (수핵난자와 전기적 융합조건이 산양의 이종간 복제수정란의 체외발달에 미치는 영향)

  • 이명열;박희성
    • Reproductive and Developmental Biology
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    • v.28 no.1
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    • pp.21-27
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    • 2004
  • This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS + 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P<0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P<0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).

The Effects of Calcium-Phosphate Coated Xenogenic Bone and Type I Collagen for Bone Regeneration on the Calvarial Defects in Rabbits (Ca-P 박막 이종골과 제 1형 교원질이 토끼 두개골 결손부의 골재생에 미치는 영향)

  • Kim, Chang-Han;Park, Jin-Woo;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.34 no.1
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    • pp.223-241
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    • 2004
  • The purpose of this present study evaluated the osseous response around Ca-P coated xenogenic bone and compared osteogenic potential of Ca-P coated xenogenic bone to that of combination with type I collagen derived from bovine tendon as a biocompatible binder to prevent migration of bone particle on the repair of calvarial defects in rabbits. To study the effects of Ca-P coated xenogenic bone and collagen on bone healing, four 5-mm-diameter skull defect were made in calvaria with trephine filled with an autogenous bone chip or Ca-P coated xenogenic bone or Ca-P coated xenogenic bone and type I collagen (1:1 mixture by volume) or left empty. The defects were evaluated histologically at 1, 2, 4 and 8 weeks following implantation. Ca-P coated xenogenic bone at the calvarial defects of rabbits showed osteoconductivity at the margin of defect in the early stage of bony healing, but no direct contact with new bone was observed. With time passed by, it was resorbed slowly and showed consistent inflammatory reaction. An additional use of type I collagen derived from bovine tendon improved clinical handling, but no new bone formation was observed histologically. Above all, autogenous bone graft showed most prominent healing in quantity and density of new bone formation. According to this study, the use of Ca-P coated xenogenic bone alone and combination with type I collagen did not showed effective healing in quantity and density of new bone formation.

The Effect of Transplantation of Schwann Cell and SIS Sponge on the Injured Peripheral Nerve Regeneration (슈반세포와 SIS 스폰지의 이식이 손상된 말초 신경 재생에 미치는 영향)

  • Kim, Cho-Min;Kim, Soon-Hee;Kim, Su-Mi;Park, Sang-Wook;Lee, Il-Woo;Kim, Moon-Suk;Rhee, John-M.;Khang, Gil-Son;Lee, Hai-Bang
    • Polymer(Korea)
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    • v.32 no.1
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    • pp.49-55
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    • 2008
  • It is recognized that Schwann cells (SC) are essential for peripheral nerve development and regeneration. SIS (small intestinal submucosa) consists of some growth factors which can stimulate cell activity without immune rejection responges. SCs were harvested from the femurs and tibias of female Fischer rat and then suspended with $2{\times}10^6$ cell/sponge in SIS sponge. Fischer rat received an implant consisting of the SCs and the SIS sponge at the place of a 5 mm gap created by the sciatic nerve resection. Thin sections were stained with H &E staining and immunostaining of S-100, GFAP and NF after 1, 2, and 4 weeks. It was observed that the effects of the SIS sponge with SCs on neuroinduction(Group II, with scaffold & cell) are strong as much as uninjured model(Control I), and significantly stronger than SIS sponge model (Group 1, with scaffold only) and blank model (Control II). In conclusion, these results suggest that SIS sponge filled with SCs may have an important role for peripheral nerve regeneration of tissue engineering.

Aquatitative analysis by digital subtraction radiography on the effect of Enamel Matrix Protein (Emdogain) and Platelet Rich Plasma, combined with xenograft in the treatment of intrabony defect in humans (수직성 골내낭 치료 시 법랑 기질 단백질과 이종골 이식 및 혈소판 농축 혈장의 골 재생에 대한 디지털 공제술을 통한 정량적 분석)

  • Han, Geum-A;Im, Seong-Bin;Jeong, Jin-Hyeong;Hong, Gi-Seok
    • 대한치주과학회:학술대회논문집
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    • 2004.11a
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    • pp.119-120
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    • 2004
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