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Generation of Heterozygous Gal KO Fibroblast Expressing Efficiently MCP at GalT Locus
Keon Bong Oh,Nayouna Ko,Bella Kim,Seong-soo Hwang,Jeong Woong Lee,Dong-Hoon Kim,Gi-Sun Im,Byoung-Chul Yang,Man-Jong Kang,Choon-Keun Park,Jin-Ki Park 한국동물번식학회 2011 Reproductive & Developmental Biology(Supplement) Vol.35 No.2s
Generation of Heterozygous Gal KO Fibroblast Expressing Efficiently MCP at GalT Locus
Keon Bong Oh,Nayouna Ko,Bella Kim,Seong-soo Hwang,Jeong Woong Lee,Dong-Hoon Kim,Gi-Sun Im,Byoung-Chul Yang,Man-Jong Kang,Choon-Keun Park,Jin-Ki Park 한국동물생명공학회(구 한국동물번식학회) 2011 Reproductive & developmental biology Vol.35 No.2
연부조직의 손상을 동반한 원위 경골 골절에 대한 Brooker 골수내정을 이용한 치료
김종오,이기웅,고영도,강충남,오종건,유재두,윤여헌,김동욱 대한골절학회 1999 대한골절학회지 Vol.12 No.4
The treatment of distal tibial fractures with compromised soft tissue poses many problems that usually occurs from the high-energy trauma, and the results are often unsatisfactory following lots of complications like loss of reduction, malunion, and infection. We studied to evaluate the treatment results of Brooker intramedually nailing for the distal 1/3 tibial fractures with compromised soft tissue. Twenty-three cases of distal tibial fractures with compromised soft tissue were reviewed and we analyzed the results of surgical treatment in the viewpoint of union time, loss of reduction, malunion, complication and its final outcome. The range of follow-up was 24 months to 38 months with mean 29 months follow-up. Most of patients were between twenty and sixty years, and average age was 43.2 years. Acording to Gustilo and Anderson s classification, 3 were Type I, 2 were Type II of 5 open fractures. According to Tscherne’s classification, 13 were Grade I, 5 were Grade II of 18 closed fractures. The average to union was 15 weeks with range 11 to 20 weeks. The healing was slowest in Tscherne’s Type II and fastest in Tscherne s Type I fracture. There were 3 cases of malunion, more than 5 degrees. All of the 3 cases were posterior angulation. Only 1 case was the loss of reduction. This case was 3 to 10 degrees of varus angulation. There were 3 cases of superficial infection. The infection was controlled with antibiotic therapy. Only 1 case was acceptable of the final outcome. This case was limping gait because of pain and loss of ankle dorsiflexion to 15 degrees. But, the limitation of ordinary work was not seen. And 18 cases were excellent and 4 cases were good. We recommand that wherever possible, Brooker intramedually nailing can be used for distal tibial fractures with compromised soft tissue. And a high rate of union and a low rate of complication can be expected with this treatment modality.
Seongsoo Hwang,Keon Bong Oh,Dae-Jin Kwon,Sun A Ock,Jeong-Woong Lee,Jin-Ki Park,Gi-Sun Im 한국동물번식학회 2012 Reproductive & Developmental Biology(Supplement) Vol.36 No.2s
Here we report the productions of genetically modified cloned Massachusetts General Hospital miniature pig (MGH minipig) using freshly thawed donor cells equilibrated with roscovitine. Fibroblasts were isolated from the ear skin of a 10-day-old male MGH minipig. The donor cells were divided into two groups: cultured for 3 days (culture) and freshly thawed with 500 nM roscovitine. The viability of the donor cells was significantly higher at 0 h (94.6±3.5) compared with 1 h (81.7±5.7) after thawing (p=0.028). After 1 hr of equilibration time, the proportion of G0/G1 stage in roscovitine group was not different from 0 hr group, but not in culture medium group (p<0.01), respectively. Although the developmental characteristics were not different in both methods, the pregnancy and delivery rate in freshly thawed group were significantly higher than that of culture group (p<0.01), respectively. In total, 12 TG cloned MGH minipigs were delivered and the individual cloning efficiency was from 0 to 2.54%. Taken together, the use of freshly thawed donor cells equilibrated with roscovitine may be helpful method to increase the productivity of the genetically modified cloned MGH minipigs.
Ko, Nayoung,Lee, Jeong-Woong,Hwang, Seong Soo,Kim, Bella,Ock, Sun A,Lee, Sung-Soo,Im, Gi-Sun,Kang, Man-Jong,Park, Jin-Ki,Oh, Sung Jong,Oh, Keon Bong Marcel Dekker 2013 Animal biotechnology Vol.24 No.4
<P>Xenotransplantation of pig organs into primates leads to hyperacute rejection (HAR). Functional ablation of the pig α 1,3-galactosyltransferase (GalT) gene, which abrogates expression of the Gal α 1-3Gal β 1-4GlcNAc-R (Gal) antigen, which inhibits HAR. However, antigens other than Gal may induce immunological rejection by their cognate antibody responses. Ultimately, overexpression of complement regulatory proteins reduces acute humoral rejection by non-Gal antibodies when GalT is ablated. In this study, we developed a vector-based strategy for ablation of GalT function and concurrent expression of membrane cofactor protein (MCP, CD46). We constructed an MCP expression cassette (designated as MCP-IRESneo) and inserted between the left and the right homologous arms to target exon 9 of the GalT gene. Nucleofection of porcine ear skin fibroblasts using the U-023 and V-013 programs resulted in high transfection efficiency and cell survival. We identified 28 clones in which the MCP-IRESneo vector had been successfully targeted to exon 9 of the GalT gene. Two of those clones, with apparent morphologically mitotic fibroblast features were selected through long-term culture. GalT gene expression was downregulated in these 2 clones. Importantly, MCP was shown to be efficiently expressed at the cell surface and to efficiently protect cell lysis against normal human complement serum attack in vitro.</P>